6-(alpha, alpha, alpha-trisubstituted-acetamido) penicillanic acids and salts thereof



United States Patent 3,245,983 G-(ot,a,ot-TRISUBSTITUTED-ACETAMIDO)PENICIL- LANIC ACIDS AND SALTS THEREOF Frank Peter Doyle, Betchworth, and John Herbert Charles Nayler, Catford, London, England, assignors to Beecham Group Limited, Brentford, England, a British company No Drawing. Continuation of applications Ser. No. 858,337, Dec. 9, 1959, Ser. No. 3,479, Jan. 20, 1960, and Ser. Nos. 3,502-3,510, Jan. 20, 1960. This application Aug. 28, 1961, Ser. No. 134,120

Claims priority, application Great Britain, Jan. 22, 1959, 2,435/59; Oct. 22, 1959, 35,546/59; Oct. 26, 1959, 36,204/59; Nov. 11, 1959, 38,304/59 13 Claims. (Cl. 260239.1)

This application is a continuation of our prior, copending applications, Serial Number 858,337, filed December 9, 1959 and Serial Numbers 3,479, 3,502, 3,503, 3,504, 3,505, 3,506, 3,507, 3,508, 3,509 and 3,510 all filed January 20, 1960, and all now abandoned.

This invent-ion relates to new synthetic compounds of value as antibacterial agents, as nutritional supplements in animal feeds, as agents for the treatment of mastitis in cattle and as therapeutic agents in poultry and animals, including man, in the treatment especially of infectious diseases caused by Gram-positive bacteria and, more particularly, relates to 6-(u,a,a-trisubstituted acetamido) penicillanic acids and nontoxic salts thereof.

G-aminopenicillanic acid has the structural formula:

and is prepared according to Batchelor et al. (Nature, 183, 257-258, January 24, 1959), or Belgian Patent 569,728.

It has now been found according to this invention that a particularly useful class of derivatives is obtained by the reaction with 6-aminopenicillanic acid of an acid chloride, bromide, anhydride or mixed anhydride derived from a tri-substituted acetic acid.

Antibacterial agents such as benzylpenicillin have proved highly effective in the past in the therapy of infections due to Gram-positive bacteria but suffer from the drawback of being ineffective against numerous so-called re sistant strains of bacteria, e.g., penicillin-resistant strains of Staphylococcus aureus (Micrococcus pyOgenes var. aureus) which produce penicillinase. Many of the compounds of the present invention, in addition to their potent antibacterial activity, exhibit resistance to destruction by acid or by penicillinase or are effective against benzylpenicillin-resistantstrains of bacteria or inhibit penicillinase and thus potentiate the action of benzylpenicillin when admixed therewith and are safe 'for use in patients who cannot be given benzylpenicillin because they exhibit allergic reactions thereto.

The present invention therefore provides new penicillins having the structural formula:

Where R and R are the same or different and are each a halogen atom or an alkyl, alkenyl, aryl, aralkyl, cycloalkyl or heterocyclic group which groups may be substituted, or R and R together with the carbon atom to which they are attached form an aromatic, cycloaliphatic, or heterocyclic ring system, and R is any one of the groups defined under R and R or the group CR or SR Where R is an alkyl, alkenyl, aryl, aralkyl or cycloalkyl group and nontoxic salts thereof.

The salts are nontoxic salts including nontoxic metallic salts such as sodium, potassium, calcium and aluminum, the ammonium salt and substituted ammonium salts, e.g., salts of such nontoxic amines as trialkylamines, including triethylamine, procaine, dibeniylamine, N- benzyl-beta-phenethylamine, l-ephenamine, N,N-dibenzyl-ethylenediamine, dehydroabietylamine, N,N-bis-dehydroabietylethylenediamine, and other amines which have been used to form salts with benzylpenicillin. Also included within the scope of the present invention are easily hydrolyzed 'esters which are converted to the free acid form by chemical or enzymatic hydrolysis.

One preferred class of new penicillins according to the present invention hasthe structural Formula II above in which R is a substituted aromatic ring, at least one of the substituents being a hydroxy group, and R and R are the same or different and are each R or an alkyl, aryl or aralkyl group which may be substituted.

A further class has the structural Formula II in which R is a substituted or unsubstituted benzene ring, R is an alkyl group or an allg'oxy group each of which group contains from 1 to 6 carbon atoms inclusive, and R is any one of the groups defined under R and R A still further class has the structural formula:

where X, Y and Z are the same or different and are each hydrogen, nitro, or a halogen atom or an amino, alkylamino, dia lkyla'mino, acylamino, alkanoyl'amino, alkyl, alkoxy, hydroXy, sulphamyl, benzyl, cyclohexyl, cyclopentyl or trifluoromethyl groups and A and B are the same or different and are each an alkyl, phenyl or phenylalkyl group.

The present invention further provides a process for the preparation of new pen-icillins having the general Formula II in which '6-aminopenicillanicacid, or the fermentation liquor containing 6-aminopenicillanic acid obtained by growing a penicillin-producing mould in a nutrient medium with or without subsequent partial purification is reacted with an acid chloride, bromide, anhydride or mixed anhydride (with other carboxylic acids, including monoesters, and particularly lower aliphatic esters, of carbonic acid) derived from a trisubstituted acetic acid of the general formula:

Rl R CCOOH where R R and R are as defined above.

The following are examples of carboxylic acids of the general Formula IV [the abbreviations Ph, Me and Et are used herein for the phenyl, methyl and ethyl groups respectively]- (a) Triaryl type:

PhaC.COOH rhroooorr Pmocoon Me Me PIMILCOOH rmt looon (H0-)3.C-C OOH I I SMe SOzMe I P11000011 v l P11 00001-1 2 11 11 Me 11 R R 11 Me Lie Me 11 011m, 01111 19, Me '11 01 v 11 N0,

(b) D1'arylaIkyl type:

1 11 :.00011 mac-00011 11000011 CH; CHJCH: CH;(CH): P11100001: P11100001: P1110.c00P1 0111(0119, 'PhCH:

(c) Aryldialkyl type: 01191000011 011 0119100001! 011,011,01191000011 Ph Ph Ph 40 I '0H,0H1011,0H=)10.000r1 v I 0.00011 Ph II I Ph (d) Trlalkyl type:

011 11000011 (CH;CH1);C.CO0H cnacgzcnmoc 0 011 (cmcmcmcnmceoou (e) Diaryl (substituted oxy) type:

- rr' P11000011 12111000011 Ph1C.CO0II OCH; 0011,0111 0(011910111 P11,0.00011 P110.00011 P11000011 0011(0111), OPh 0011,1 11 (f) Diaryl (substituted thio) type:

P11,0.c0011 P11,( :.00011 P1110.00011 semen, 80311910111 s(CH1 10H1 Ih;C.COOH P10000011 P11, 3.00011 scH(0H1): t e

SPl1 IS l Me 4 (g) Dialkyl (substituted oxy) type:

(CH3)2C.COOH (C HJCIIzCHz)zC.COOII Ph 0 Pil (It) Diul kyl (substituted tlu'o) type:

(011.3)2C-COO1I (CU3CI'IZ)2C.COO1[ S Ph SP]! (CH;CH2CH2)ZC .C O H:

s'ph

(CH3CH2CII2):C.COOII SCI-ECU:

(i) Arylalkyl (substituted oxy or thio) type:

P11 CH:CHzCH2( .COOH

OCHzCIIa (j) Diaryl heterocyclic type:

PhflilCOOIlI Ph2C.COOH PhzC.COOII PhzC.COOII Mc-U;\le (k) l-arylcyeloalkyl type:

00011 00011 3 4 Ph P11 (1) 9-substituted 9-fluorenyl type:

(m) Miscellaneous:

COOII R=II or Me CI;C.COOI1 (lJ.COOH (CHaCIIzMKIZCOOII Cl Br In those cases where the acid contains a grouping such as amino or alkylamino which might react with an acylating agent, such a group may be protected in the conventional manner prior to formation of the acid chloride, bromide, anhydride or mixed anhydride. The subsequent removal of the protecting group to form the free aminosubstituted or alkylamino-subst-ituted penicillin ca be effected by catalytic hydrogenation, e.g., with palladium or platinum 0n barium carbonate or on carbon. Suitable protecting groups are the trityl group and groups of the general formula R--O-CO wherein R is an allyl, benzyl, substituted benzyl, phenyl or substituted phenyl.

Alternatively, those penicillins wherein the substituent is an amino group may be prepared by forming the corresponding nitro penicillin which is then hydrogenated by conventional means to form the amino derivative.

When an acid chloride, acid bromide or acid anhydride is used in a process of the present invention, it is prepared from the corresponding trisubstituted acetic acid according to the techniques set forth in the chemical literature, e.g., for acids such as diphenylacetic acid or as described in examples below. In instances where the trisubstituted acetic acid has not been described, it is prepared by the methods described in the chemical literature for simpler members of the series or by methods described or referred to in the examples below or by the methods described or referred to in The Chemistry of the Carbon Compounds, Richter-Auschutz, Third English Edition, vol. I, pp. 296-309, 419, and Vol. III, pp. 555-558 (1946) Elsevier Publishing Co-., 'Inc., New York, New York, or in Chemistry of Carbon Compounds, E. H. Rodd, Editor, vol. I, pp. 573-579, 788 et sub, vol. II, pp. 110-112, 220-224, and vol. HI, pp. 434, 593-601, 1127-1134, 1202-1203 (1952), Elsevier Publishing Co., Inc, New York, New York, or in Organic Preparations by Weygand, pp. 159-162, 197, 299-300 (1945), Interscience Publishers Inc, New York, New York, especially with regard to hydrolysis to the desired acid of the correspondin-g amide or nitrile or in United States Patents 2,854,450 and 2,479,297 or in J. Amer. Chem. Soc., 70, 1153 (1948), or in Acta. Chem. Scand., 7, 596-602 (1953).

One method for preparing a compound of the present invention by way of mixed anhydride with ethoxy or isobutoxy-carbonic acid comprises mixing an acid of the general Formula IV with isobutyl chloroformate and a tertiary hydrocarbonyl or aliphatic amine such as triethylamine in an anhydrous, inert and preferably water-miscible solvent such as p-dioxane and, if desired, pure dry acetone, for about thirty minutes in the cold. To this solution of the mixed anhydride there is then added a chilled solution of G-aminopenicillanic acid and tertiary hydrocarbonyl amine, e.g., triethylamine in a solvent such as water. The reaction mixture is stirred for a period of an hour or so to form the substituted ammonium salt of the desired product. The mixture may then, if desired, be extracted at alkaline pH (such as pH 8; aqueous sodium bicarbonate may be used, for example, if necessary to adjust the pH) with a water-immiscible solvent such as ether to remove unreacted starting materials. The product in the aqueous phase is then converted to the free acid, preferably in the cold under a layer of ether by the additon of dilute mineral acid, e.g., 5 N H 80 to pH 2. The free acid is then extracted into a water-immiscible, neutral organic solvent such as ether and the extract is washed with water quickly in the cold, if desired, and then dried, as with anhydrous Na SO The product in the ethereal extract in its free acid form is then converted to any desired metal or amine salt by treatment with the appropriate base, e.g., a free amine such as procaine base or a solution of potassium 2-ethylhexanoate in dry nbutanol. These salts are usually insoluble in solvents such as ether and can be recovered in pure form by simple filtration.

Another method of preparing an ethereal solution of the acid form of a compound of the present invention comprises preparing an aqueous solution of 6-aminopenicil-lanic acid and sodium bicarbonate, adding an acid chloride derived from an acid of the general Formula IV and shaking vigorously at room temperature, e.g., for twenty to sixty minutes. The mixture is then extracted with ether to remove unreacted or hydrolyzed starting materials. The solution is then acidified (preferably in the cold) to pH 2, as with dilute sulphuric acid, and the free acid form of the product is extracted into ether. This ethereal extract is dried, as with anhydrous sodium sulphate, and the drying agent is removed to leave a dry ethereal solution from which the product is easily isolated, preferably in the form of an ether-insoluble salt such as the potassium salt. This procedure is used when the acid chloride reacts With a primary amine more rapidly than it does with water, as determined by simple test. In this procedure the acid chloride may be replaced by the corresponding acid bromide or acid anhydride.

Since some of the antibiotic substances obtained by the process of this invention are relatively unstable compounds which readily undergo chemical changes resulting in the loss of antibiotic activity, it is desirable to choose reaction conditions which are sufficiently moderate to avoid their decomposition. The reaction conditions chosen will, of course, depend largely upon the reactivity of the chemical reagent being used. In most instances, a compromise has to be made between the meet very mild conditions for a lengthy period and the use of more vigorous conditions for a shorter time with the possibility of decomposing some of the antibiotic substances.

The temperature chosen for the process of preparation of the derivatives of penicillanic acid should in general not exceed 30 C. and in many cases a suitable temperature is ambient temperature. Since the use of strongly acid or alkaline conditions in the process of this invention should be avoided, it has been found preferable to perform the process at a pH of from 6 to 9, and this can conveniently be achieved by using :a buffer, for example, -a solution of sodium bicarbonate, or a sodium phosphate buffer. In addition to the use of aqueous media for the reaction, including filtered fermentation broths or aqueous solution of crude 6-aminopenicillanic acid, use can be made of organic solvents which do not contain reactive hydrogen atoms. Examples of such inert solvents are dimethylformamide, dimethylacetamide, chloroform, acetone, methylisobutyl ketone and dioxane. requently it is highly satisfactory to add an aqueous solution of a salt of '6-aminopenicillanic acid to a solution of the acylating agent in an inert solvent and preferably in an inert solvent which is miscible with water, such as acetone or dimethylformamide. Vigorous stirring is of course advisable when more than one phase is present, e.g., solid and liquid or two liquid phases.

At the conclusion of the reaction, the products are isolated if desired by the techniques used with benzylpenicillin and phenoxymethylpenicillin. Thus, the product can be extracted into diethyl ether or n-butanol at acid pH and then recovered by lyophilization or by conversion to a solvent-insoluble salt, as by neutralization with an n-butanol solution of potassium 2-ethylhexanoate, or the product can be precipitated from aqueous solution as a water-insoluble salt of an amine or recovered directly by lyophilization, preferably in the form of a sodium or potassium salt. When formed as the triethylamine salt, the product is converted to the free acid and thence to other salts in the manner used with benzylpenicillin and other penicillins. Thus, treatment of such a triethylamine compound in water with sodium hydroxide converts it to the sodium salt and the triethylamine may be removed by extraction, as with toluene. Treatment of the sodium salt wi-th strong aqueous acid converts the compound to the acid form, which can be converted to other amine salts, e.g., procaine, by reaction with the amine base. Salts so formed are isolated by lyophilization or, if the product is insoluble, by filtration. A particularly elegant method of isolating the product as a crystalline potassium salt comprises extracting the product from an acidic, aqueous solution (e.g., pH 2) into diethyl ether, drying the ether and adding at least one equivalent of a solution of potassium Z-ethyIheX-anoate in dry n-butanol. The potassium salt forms, precipitates, usually in crystalline form, and is collected by filtration or decantation.

It is sometimes preferred to prepare the new penicillins from the original fermentation liquor, containing 6-aminopenicillanic acid or one of its intermediate concentrates.

Some of the new penicillins of the present invention have shown a particular advantage in that they are less readily destroyed by penicillinase than the conventional penicillins G and V. The following table shows the relative activity of one of these new compounds as compared with penicillin G when tested against three hospital-resistant strains of staphylococci in vitro.

' Minimum Inhibitory Concentration Compound Resistant Resistant Resistant Strain 1 Strain 2 Strain 3 Ph.CHz.CO.NH.- 114,000 1:400, 000 1:1, 000, 000

(250 n1cg./ (2.5 mcg./ (1 meg. ml.) ml.) ml.) C|}HCH O.Me2 G N CH. C 0211 Penicillin G Ph3.C.CO.NH. 1:3, 200, 000 1:6, 400, 000 1:6, 400, 000 (0.3 mag] (0.16 mcg./ (0.16 meg] ml.) 1111-) ?IICI-I C .Mez

C ONCH.C 0 H (a) TRIARYL TYPE Example 1 .-Trz'phenylmethylpenicillin (1') Crude sodium salt.-A solution of triphenylacetyl chloride (0.8 g.) in chloroform ('10 ml.) was added during 5 minutes to a suspension of 6-aminopenicillanic acid 0.5 gm. in chloroform (20 ml.) and triethylamine (1 ml.). After stirring at room temperature for 1 hour the solution was filtered and the solvent evaporated in vacuo at low temperature. The semi-solid residue was stirred in n-butanol (50 ml.) and water (30 ml.), was added, followed by sufiicient N. hydrochloric acid to reduce the pH of the aqueous phase to 2.0. After shaking, the aqueous layer was discarded and the butanol washed twice further with very dilute hydrochloric acid. The penicillin was obtained from the butanol layer by shaking it with suflicient sodium bicarbonate solution to give a neutral aqueous phase (pH 7), separating the latter, and removing water at low temperature and pressure to leave the crude sodium salt of triphenylmethyl penicillin as a pale yellow solid (0.9 g.). It inhibited Staph. aureus at a concentration of 0.3 meg/m1, the benzylpenicillin-resistant Staph. 1. at 0.6 meg/ml, and the benzylpenicillin-resistant Staph. 2. at 0.6 meg/ml.

(ii) Benzylamine salt.The crude sodium salt of triphenylmethylpenicillin (150 mg.) prepared according to (i) was suspended in water (15 ml.) covered with an equal volume of ether, and acidified to pH 2 with dilute hydrochloric acid so that upon shaking, the free pencillin acid passed into the ether layer. This ether solution was separated, washed with water, dried over anhydrous sodium sulphate, filtered, and treated with 2 ml. of a 2% solution of benzylamine in ether. The benzylamine salt of the penicillin was precipitated as an oil which slowly solidified on trituration, probably taking up Water of crystallization from the atmosphere in the process. The

resulting white powder was collected, washed with ether,

and dried in a vacuum desiccator. Yield 65 mg., M.P. 98100 C. (decomp). (Found: C, 66.9; H, 6.2; N, 6.8; S, 4.5. C H O N S, 2H O requires: C, 66.7; H, 6.2; N, 6.7; S, 5.1%.)

(iii) Cyclohexylamine salt.-A solution of of triphenylmethylpenicillin in ether was prepared from the crude sodium salt (180 mg.) as in (ii). Treatment with 2 ml. of a 2% solution of cyclohexylamine in ether precipitated the cyclohexylamine salt of the penicillin as a slightly deliquescent white powder of indefinite M.P. which was then washed with ether (2 x 600 ml.).

itated the N-methylmorpholine salt of the penicillin as a colourless oil which slowly solidified when rubbed. The resulting white powder was collected, washed with ether, and dried in a vacuum desiccator. Yield 60 mg., M.P. 118121 C. (decomp). (Found: C, 66.6; H, 6.3; N, 6.9; S, 5.3. C H O N S, 0.5H O requires: C, 66.4; H, 6.4; N, 7.0; S, 5.4%.)

(v) Pure sodium salt.-A solution of pure triphenylacetyl chloride (18.4 g.) in dry acetone (360 ml.) was added during 15 mins. to a stirred solution of 6-aminopenicillanic acid (13 g.) in a mixture of 3% aqueous sodium bicarbonate solution (500 ml.) and acetone (150 ml.). The mixture was stirred at room temperature for 3 hrs., The aqueous layer was filtered to remove a little suspended solid and then cooled to 6 C., whereupon the product crystallized in glistening plates. It was collected (refrigerated centrifuge) and recrystallized by dissolution in aqueous acetone (300 ml.) at room temperature followed by cooling to -6 C. The pure crystalline sodium salt was collected as before and dried in a vacuum desiccator to give a white powder (17.1 g., M.P. 120 C.) which appeared to be essentially the monohydrate. Further drying over phosphorus pentoxide at 80 C./0.4 mm., gave the anhydrous salt, M.P. 183190 C. (decomp.), but this was hygroscopic and on exposure to air the melting point quickly fell to 100-120 C.

A specimen for analysis was recrystallized and dried once more as described above. (Found: C, 65.9; H, 5.2; N, 5.2; S, 6.2; Na, 4.4. C H O N SNa required: C, 66.1; H, 5.0; N, 5.5; S, 6.4; Na, 4.5%.) [u] +8l C. (1.5% in water).

(vi) Pure potassium salt.This was prepared exactly as described in (v) above except that the equivalent quantity of potassium bicarbonate was substituted for sodium bicarbonate. After being isolated and dried as described for the sodium salt the anhydrous potassium salt had M.P. 199-201 C. (decomp). (Found: C, 63.7; H, 4.8; N, 5.1; S, 5.8; K, 7.4. C H O N SK required: C, 64.1; H, 4.8; N, 5.3; S, 6.1; K, 7.4%.)

Example 2.M0ua-p-hydr0xytriphenylmethylpenicillin Monophydroxytriphenylacetyl chloride (7.2 g.) was dissolved in dry acetone and added over about 7 minutes to a stirred solution of G-aminopenicillanic acid (4.84 g.) in a mixture of aqueous sodium bicarbonate ml. of 8.8%) and acetone (75 ml.). The mixture was stirred at room temperature for 3 hrs., diluted with water (50 ml.), and extracted with ether (2 x 100 ml.) only the aqueous phase being retained. This aqueous solution Was covered with ether (80 ml.), acidified to pH 2 to 3 with N. hydrochloric acid (92 ml.), shaken, and the ether layer separated. The aqueous layer was extracted with two further 40 ml. portions of ether and the combined ether extracts (which at this stage contained the free penicillin acid) were washed with water (20 ml.) and then neutralized with 8.4% aqueous sodium bicarbonate (13.5 ml.), with vigorous shaking. The aqueous layer was separated and evaporated to dryness in vacuo at room temperature to give the sodium salt of mono-p-hydroxytriphenylmethyl penicillin as a yellow solid (6.8 g.).

It inhibited Staph. Oxford at a concentration of 0.5 mcg./ml., the benzylpenicillin-resistant Staph. l. at 2.5

.mcg./ml. and the benzylpenicillin-resistant Staph. 2. at .2.5 mcg./ml.

in benzene solution together with pyridine in catalytic amount. The reaction was effected by heating at 60-70 C. for 1 hour and the acid chloride isolated by concentrating the reaction mixture. Its purity was assessed by model reactions with aniline and ammonia in presence of water to give the anilide and amide respectively.

Example 3.Mon-p-mcth-0xytriplzenylmefhylpenicillin A solution of mono-p-methoxytriphenylacetyl chloride (673 mg.) in dry chloroform ml.) was added dropwise to a stirred mixture of 6-aminopenicillanic acid (432 mg.), chloroform ml.), and triethylamine (0.84 ml.). The mixture was stirred at room temperature for 2 /2 hrs., and then filtered. The filtrate was treated with successive small portions of 0.2 N. hydrochloric acid, with vigorous shaking between additions, until the aqueous phase was at pH 2. The acid layer was separated and discarded, and the chloroform solution was washed with water ml.). Sufiicient 3% aqueous sodium bicarbonate solution was added to the chloroform solution to give, after vigorous shaking, a neutral emulsion (pH 7). This was evaporated at low temperature and pressure and the residual hygroscopic solid dried in a vacuum desiccator. There was obtained 817 mg. of the sodium salt of monop-methoxytriphenylmethylpenicillin.

The product inhibited Staph. anrens at a concentration of 0.5 mcg./ml., the benzylpenicillin-resistant Staph. l. at 0.6 mcg./ ml. and the benzylpencillin-resistant Staph. 2. at 0.6 meg/ml.

The mono-p-methoxytriphenylacetyl chloride was prepared from the corresponding acid by heating with thionyl chloride and a trace of pyridine in benzene, removing the solvent and excess reagent in vacuo and crystallizing the residue from light petroleum, M.P. 101l03 C.

Example 4.M0no-p-mezhylthiotriphenylmethylpenicillin A solution of mono-p-pmethylthiotriphenylacetyl chloride (6.34 g.) in dry acetone (108 ml.) was added slowly to a stirred solution of 6-aminopenicillanic acid (3.89 g.) in 3% aqueous sodium bicarbonate (152 ml.) and acetone (45 ml.). The mixture was stirred at room temperature for 4 hrs., then washed with two 200 ml. portions of ether. The aqueous phase was covered with ether (50 ml.) and adjusted to pH 2 "by the addition of N. hydrochloric acid. After separating the layers, the aqueous phase was extracted with two further 50 ml. portions of ether. The combined ether solutions (which at this stage contained the free penicillin acid) were washed with water and then shaken with sufficient 8% sodium bicarhonate solution to give a neutral aqeuous phase (pH 7). The layers were separated and the aqueous phase was evaporated at low temperature and pressure to leave the sodium salt of mono-p-methylthiotriphenylmethylpenicillin as a white solid, which was finally dried in a vacuum desiccator. Yield 7.44 g.

The product inhibited Staph. Ox-ford at a concentration of 0.025 meg/ml, Staph. 1. at 0.25 mcg./ml., and Staph. 2. at 1.25 meg/ml.

The monop-methylthiotriphenylacetyl chloride was prepared from the corresponding acid by heating with thionyl chloride and a trace of pyridine in benzene, removing the solvent and excess reagent in vacuo and crystallizing the residue from light petroleum, M.P. 103.5-105 C.

Example 5 .M0no-p-methylsulphonylzriplzenylmethylpenicillin G-aminopenicillanic acid (4.32 g.) was acylated with mono-p-methylsulphonyltriphenylacetyl .chloride (7.68 g.) as described in Example 4 to give the sodium salt of mono-p-methylsulphonyl-triphenylmethylpenicillin as an almost whitesolid (9.23 g.).

It inhibited Staph. Oxford at a concentration of 0.6

10 meg/1111., Staph 1. at 2.5 mcg./ml., and Staph 2. at 2.5 mcg./ml.

The mono-p-methylsulphonyltriphenylacetyl chloride was prepared from the corresponding acid by heating with thionyl chloride and a trace of pyridine in benzene, removing the solvent and excess reagent in vacuo, and crystallizing the residue from light petroleum, M.P. 146- 148 C.

Example 6.u-(4-methyl-1-naphthyl)diphenylmetlzylpenicillin 6-aminopenicillanic acid (4.85 g.) was acylated with ow(4-methyl-l-naphthyl)diphenylacetyl chloride (8.3 g.) as described in Example 3 to give the sodium salt of tit-(4- methyl-l-naphthyl)diphenylmethylpenicillin as a white solid (9.7 g.).

It inhibited Staph. Oxford at a concentration of 12.5 mcg./ml., Staph. 1. at 25 meg/ml, and Staph. 2. at 25 meg/ml.

The a-(4-inethyl-l-naphthyl)diphenylacetyl chloride M.P. 176'177 C. (decomp.), was prepared by heating the corresponding acid in benzene with thionyl chloride and a trace of pyridine.

Example 7 .Tri-( p-methoxyphenyl methyl penicil lin 6-aminopeni-c-i1lanic acid (2.94 g.) was acylated with crude tri-(p-methoxyphenyl)acetyl chloride (obtained by heating 5.14 g. of the corresponding acid in benzene with thionyl chloride, then removing the solvent and excess reagent in vacuo) according to the method described in Example 4, except that the solvent extractions were carried out with butanol instead of ether. There was obtained 3.44 g. of sodium salt of tri-(p-methoxy-phenyl) methylpenicillin as a slightly hygroscopic white powder.

It inhibited Staph. Oxford at a concentration of 5 meg/1111., Staph. 1. at 12.5 mcg./ml., and Staph. 2. at 12.5 meg/ml.

Tri fip-methoxyphenyl)acetic acid, M.P. 212-213 C., was prepared in good yield by condensing anisilic acid with an-isole in the presence of sulphuric and acetic acids.

Example 8 In the procedure of Example 2 the mono-p-hydroxytriphenylaoetyl chloride is replaced by an equimolar weight of the acid chloride of a-(4-chlo-rophenyl)-u,u-diphenylacetic acid, u-'(3-bromophenyl)-a,a-diphenylacetic acid,

a (2-iodophenyl)-t,u-diphenylacetic acid, a-(2-aminophenyl)-a,m-diphenylacetic acid, a-(3-fluorophenyl)-a,a-dihpenylacetic acid,

a- (4-nitrophenyl)-a,a-d-iphenylacetic acid,

w (4-dimethylaminophenyl)-a,u-diphenylacetic acid, a- 3-et hyl-aminophenyl -a,a-diphenyl acetic acid,

w (Z-acetamidophenyl -Ot,OL-dlphIly1a.C6tlC acid, t-(3,4-dirneithylphenyl)-a,ot-diphenylacetic acid, u-(3,4-d-ichlorophenyl)-a,a-diphenyl ace tic acid,

' a-(2benzylphenyl)-a,a-diphenylacetic acid,

a-(3-cyclohexylphenyl)-tx,a-diphenylacetic acid,

a-(4-sulfamylphenyD-u,a-diphenylacetic acid,

a-(4-trifluoromethylphenyl)-ot, x-diphenylacetic acid,

w(3,4,5-trimethoxyphenyl)-u,a-diphenylacetic acid,

tit-(2,4-dinitrophenyD-u,c-diphenylacetic acid respectively to produce the acids 6-[a-(4-chlorophenyl)-a,ot-diphenylacetamido]- penicillanic acid,

6-[a-(3-bromophenyl)-u,a-diphenylacetamido} penicillanic acid,

6- [a- (2-iodophenyl -a,zx-d-iphenylacetamido] penicillanic acid,

6- [oc- (Z-aminophenyl -a,a-diphenylacetamido penicillanic acid,

6- [oc- 3-fluorophenyl -a,a-dipheny1acetamido] penicillanic acid,

6- [a- (4-nitrophenyl) -u,ot-diphenylacetamido] penicillanic acid,

6- [a- (4-di-methyl aminophenyl) -a,u-diphenylacetamid01penicillanic acid,

6- [oc- 3 -ethylaminophenyl -u,a-dipheny1- acetamido1penicillanic acid,

6-[ oc- (Z-acetamidophenyl -a,a-diphenylacetamido1penicillanic acid,

6- oc- (3 ,4-dimethylphenyl) -a,a-diphenylacetamido] penicillanic acid,

6- OL- 3 ,4-dichlorphenyl -a,ot-diphenylacetamido] penicillanic acid,

6- [oc- (Z-benzylphenyl) a,u-dipheny1acetamido] penicillanic acid,

6- a- (3-cyclohexyphenyl) -a,a-diphenylacetamido] pencillan'ic acid,

6- a-(4-sulfamylphenyl -a,a-diphenylacetamido] penicillanic acid,

6- a- 4-trifluoromethylphenyl) -a,o-diphenyl acetamido] pencillanic acid, 6-[ix-(3,4,5-trimethoxyphenyl)-a,e-dipbenylacetamido] penicillanic acid, 6-[a-(2,4-dinitrophenyl)-a,a-diphenylacetamido] penicillanic acid, respectively, which are isolated as their solid, watersoluble sodium salts and found to inhibit Staph. aureus Smith at concentration of 0.001 percent by weight.

Example 9 In the procedure of Example 2, the mono-p-hydroxytriphenylacetylchloride is replaced by an equimolar weight of the acid chloride of u- (3 -ch-lorophenyl -a- (4 hydroxyphenyl -a-phenylacetic acid,

u- Z-bromophenyl) x,a-di (4-hydroxyphenyl) acetic acid,

04- (4-iodophenyl -oc l-methoxyphenyl) -a-phenylacetic acid,

a-(2-aminophenyl -ot,oc-di- (Z-methoxyphenyl) acetic acid,

a-(4-nitrophenyl) -a,u-di-(4-methylphenyl) acetic acid,

Ot- (4-dimethylaminophenyl -a- (Z-methylphenyl) -aphenylacetic acid,

a- 3 ,4-dichlorophenyl) w nt-di-(Z-hydroxyphenyl) acetic acid,

a,a,a-tri-(A-methylphenyl) acetic acid and a,u,u-tri(3,4-dichlorophneyl) acetic acid,

respectively to produce the acid 6- [a- 3-chl0rophenyl) -oc- (4-hydroxypheny1) -aphenylacetamido1penicillanic acid,

6- oz- (2-bromophenyl-o a-di- (4-'hydroxyphenyl) acetamido1penicillanic acid,

6- oc- (4-iodophenyl) -a-(4-methoxyphenyl) -a-phenylacetamidolpenicillanic acid,

6- a-(Z-aminophenyl) -cc,ot-di (Z-methoxyphenyl) acetamidoJpenicillanic acid,

6- a- (4-nitrophenyl -a,a-di (4-methylphenyl) acetam-ido] penicillanic acid,

6- u-(4-dimethylaminophenyl) -a- (Z-methylphenyl) a-phenylacetamido]penicillanic acid,

6- a- 3 ,4-dichlor-ophenyl) -a,u-di- Z-hydroxyphenyl) acetamido]penicillanic acid,

6-[a,a,u-tri-(4-methylphenyl) a-cetamido] penicillanic acid,

6- [a,u,a-tri (3,4-dichlorophenyl) acetamido1penicillanic acid,

respectively, which are isolated as their solid, watersoluble sodium salts and found to inhibit Staph. aureus Smith at concentrations of 0.001 percent by weight.

(b) DIARYLALKYL TYPE Example 10.a,a-a'iphenylethylpenicillin A solution of u,a-diphenylpropionyl chloride (0.49 g.) in dry chloroform (6 ml.) was added dropwise to a stirred mixture of 6-amindpenicillanic acid (0.43 g. chloroform (12 ml.), and triethylamine (0.84 ml.) Stirring was continued for 2 /2 hours and the mixture was then filtered to remove any solid which had failed to dissolve. The

filtrate was treated with successive small portions of 0.2 N. hydrochloric acid, with vigorous shaking between ad- .ditions, until the aqueous phase was at pH 2-3. The acid mcg./ml., the benzylpenicillin-resistant Staph. 1. at 6 meg/ml. and the benzylpenicillin-resistant Staph. 2. at 5 meg/ml.

Example 11 .-a,u-diphenylpropylpenicillin a,oc-diphenylbutyryl chloride (3.8 g.) in dry acetone (20 ml.) was added dropwise during 15 mins. to a stirred solution of G-aminopenicillanic acid (3.2 g.) and sodium bicarbonate (3.7 g.) in water (60 ml.) and acetone (40 ml.). When addition was complete the mixture was stirred at room temperature for 3 hrs. then diluted with water (50 ml.) and extracted with two 60 ml. portions of ether, only the aqueous phase being retained. This aqueous solution was covered with ether (50 ml.) and adjusted to pH 2-3 by the addition of N. hydrochloric acid. After separating the layers, the aqueous phase was extracted with two 25 ml. portions of ether, adjusting to pH 2-3 each time. The combined ether solutions (which at this stage contained the free penicillin acid) were washed with water (50 ml.) and then shaken with sufiicient 8% sodium bicarbonate solution to give a neutral aqueous phase (pH 7). The layers were separated and the ether phase was extracted with two 5 m1. portions of water to each of which was added sufiicient bicarbonate solution to produce an aqueous phase of pH 7. Evaporation of the combined aqueous solutions at low temperature and pressure left the sodium salt of oaadiphenylpropylpenicillin, which was finally dried in .a vacuum desiccator. Yield 5.2 g.

It inhibited Staph. aareus at a concentration of 0.006 mcg./ml., Staph. 1. at 1.25 meg/ml, and Staph 2 at 1.25 meg/ml.

Example 12.-a,ot-diphenylbutylpenicillin 6-aminopenicillanic acid (3.6 g.) was acylated with a,a-diphenylvaleryl chloride (4.5 g.) as described in Example 1 to give the sodium salt of a,a-diphenylbutylpenicillin (6.3 g.)

It inhibited Staph. Oxford at a concentration of 0.12 meg/ml, Staph. 1. at 1.25 mcg./ml., and Staph. 2. at 0.6 meg/ml.

The a,a-diphenylvaleryl chloride, B.P. -l22 C./ 0.2 mm., was prepared by heating a,a-diphenylvaleric acid with thionyl chloride in benzene.

Example 13.--a,a-diphenylpehtylpenicillin 6 aminopenicillanicacid (2.6 g.) was a'cylated with ot-diphenylhexoyl chloride (3.5 g.) as described in Example 10 to give the sodium salt of a,a-diphenylpentyl penicillin (5.8 g.).

It inhibited Staph. Oxford at a concentration of 0.25

mcg./ml., Staph. 1. at 1.25 mcg./ml., and Staph. 2. at 0.6 mcg./ml.

p The u,oc-diphenyl'hexoyl chloride, B.P. 14-2144 C./ 0.3 mm., was prepared by heating a,a-diphenylhexoic acid with thionyl chloride in benzene. 06,0t-DlPl16I1YlhCXOlC acid (needles from aqueous ethanol, M.P. 131 C.) was itself obtained by alkylating diphenylacetonitrile with butyl bromide in the presence of sodamide, and hydrolyzing the resulting a,a-diphenylhexonitrile, M.P. 132 C./ 0.05 mm., with alcoholic potassium hydroxide at C. under pressure.

Example 14.-o,ot-Diphenylis0butylpenicillin 6-aminopenicillanic acid (3.25 g.) was acylated with u,a-diphenylisovaleryl chloride (4.27 g.) according to the procedure of Example 11 to give the sodium salt of oc,0cdiphenylisobutylpenicillin (2.37 g.).

It inhibited Staph. Oxford at a concentration of 0.5 rncg./ml., Staph. l. at 1 0.0 mcg./ml., and Staph. 2., at 5.0 meg/ml.

a,u-Diphenylisovaleryl chloride, M.P. 78-79 C., was prepared by the action of thionyl chloride on the corresponding acid.

Example 15 In the procedure of Example the a,a-diphenylpropionyl chloride is replaced by an equimolar weight of the acid chloride of oc- 4-nitrophenyl -u-phenyl-n-b-utyric acid,

a- (4-bromophenyl) -a (4-hydroxyphenyl) -n-valeric acid,

a- 4-t-butylphenyl -a (4-methoxyphenyl -n-propionic acid,

a- (4-trifluoromethylphenyl -a- 4-methylphenyl) nhexanoic acid,

a- 3-aminophenyl) -u-(2-hydroxyphenyl) -isovaleric acid,

a-(4sulfarnylphenyl) -a-(2-II16thOXYphG113/1) -isobutyrie acid,

a- 2-benzylphenyl) -oz- (Z-methylphenyl) -n-valeric acid,

oc- 3-methoxyphenyl) -ot-phenyl-n-decanoic acid,

a- (2-io dophenyl -a-( 3-hydroxyphenyl-n-propionic acid,

oz- 3 -diethy1aminophenyl) -oc-( 3 -methoxyphenyl) -n-valeric acid,

oc-(4-cyclohexylphenyl) -a- (3-methylphenyl) -isohexanoic acid,

oz- (4-methylaminophenyl) -a-phenyl-a-cyclobutylacetic acid,

a- (4-chlorophenyl) -a- (4-'hydroxyphenyl) -u-cyclopentylacetic acid,

11- (Z-ace-tamid op-henyl -oc- (3 -hydroxyphenyl) flat-cyclohexylacetic acid,

11- 3,4-dimethylphenyl) -a- (2-hydroxyphenyl) -oc-CYC10- heptylacetic acid,

a- 3,4-dichlorophenyl) -a-phenyl-isohexanoic acid,

a- 3 ,4,5-trimethoxyphenyl) -a-phenyl-n-propionic acid,

ct-(4-hydroxyphenyl)-a-pheny1-n-butyric acid,

oc- (3 ,4-dihydroxyphenyl) -a-phenyl-n-propionic acid,

oc- (4-methylthiopheny1) -a-phenyl-n-butyric acid,

a- 4-methylsulfonylpheny1) -a-phenyl-n-propionic acid and u- (2,5 dimethyl-4-hydroxyphenyl) -a-phenyl-n-propionic acid,

respectively, to produce the acids 6- [a- (4-nitrophenyl) -a-phenyl-n-butyramid-o] penicillanic acid,

6- a- (4-bromophenyl) -a- (4-hyd-roxyphenyl) -n-valeramido1penicillanic acid,

6- [a-(4-t-buty1pheny1) -a-(44methoxyphenyl) -n-propionamido] penicillanic acid,

6- ac- (4-trifiuoromethylphenyl -oc- (4-rnethylphenyl) -nhexanoamido1penicillanic acid,

6- oa- 3-aminophenyl) oc- 2-hydroxyphenyl -iso-valeramido] penicillanic acid,

6- [a- (4-sulfamylphenyl -a-(2-methoxyphenyl -isobutyramido]penicil1anic acid,

6- oc- (Z-benzy-l-phenyl) -a- (Z-methylphenyl) -n-valeramido] penicillanic acid,

6- a- 3-methoxyp-henyl -a-phenyl-n-decanoamido] penicillanic acid,

6- [a- (2-iodophenyl) -oz-( 3 -hydroxyphenyl) -n-propionamido]penicillanic acid,

6- [a( 3-diethy'laminopheny1) -u- 3-methoxyphenyl) -nvaleramido] penicillanic acid,

6- [a- (4-cyclohexyl-phenyl -u- 3-methylphenyl -isohexanoamido] penicillanic acid,

6- [a- (4-methy1aminophenyl) -a-phenyl-u-cyclobutylacetamido] penicillanic acid,

Cir

respectively, which are isolated as their solid, water-soluble sodium salts and found to inhibit Staph. aureus Smith at concentrations of 0.001 percent by weight.

Example 16 In the procedure of Example 10 the a,a-diphenylpropionyl chloride is replaced by an equimolar weight of the acid chloride of a,a-di-(4-hydroxyphenyl-n-propionic acid,

m,a-di- Z-hydroxyphenyl) -n-butyric acid, a,a-di-(4-methylphenyl-n-valeric acid, a,u-di-(3-methoxyphenyl)-n-hexanoic acid, a,a-di-phcnyl-n-octanoic acid, a,u-di-(3-hydroxyphenyl) isohexanoic acid, a,u-di-(4-methoxyphenyl) -n-pr0pionic acid, a,a-diphenyl-a-cyclohexylacetic acid and ot,zx-di- (4-hydroxyphenyl) -a-cyclopentylacetic acid,

respectively to produce the acids 6- a, xdi- (4-hydroxyphenyl) -n-propionamido] penicillanic acid,

6- u,a-di- (Z-hydroxyphenyl) -n-butyramido] penicillanic acid,

6- [11, w-di- (4-methylpl1enyl) -n-valeramido penicillanic acid,

6- a, oc-di- 3 methoxyphenyl -n-hexanoamido] penicillanic acid,

6- [a,a-diphenyl-n-o-ctanoamido] penicillanic acid,

6 a,oc-di- (3-hydroxyphenyl isohexanoamido penicillanic acid,

6 [0a,oc-di- (4-rnethoxyphenyl) -n-propionamid0] penicillanic acid,

6- [a,a-diphenyl-a-cyclohexylacetamido] penicillanic acid and 6- [oz,cx-Cli (4-hydroxyp-henyl -a-cyclopentylacetamido] penicillanic acid,

respectively, which are isolated as their solid, water-sol=u ble sodium :salts and found to inhibit Staph. aureus Smith at concentrations of 0.001 percent by weight.

(0) ARYL DIALKYL TYPE Example 17.u,a-Dimelhylbenzylpenicillin A solution of u,u-dimethylphenylace-tyl chloride (5.2 g.) in dry acetone (40 ml.) was added dropwise during 15 mins. to a stirred solution of 6-aminopenicillanic acid (5.6 g.) and sodium bicarbonate (6.5 g.) in water ml.) and acetone (50 ml.). When addition was complete the mixture was stirred at room temperature for 2 /2 hrs., then diluted with water (50 m1.) and extracted with ether ml. in 3 portions), only the aqueous phase being retained. This aqueous solution was covered with ether (50 m1.) and adjusted to pH 2-3 by the addition of N. hydrochloric acid. After separating the layers, the

aqueous phase was extracted with two 25 ml. portions of ether, adjusting to pH 2-3 each time. The combined ether solutions (which at this stage contained the free penicillin acid) were washed with water (2 x 50 ml.) and then shaken with sufficient 8% sodium bicarbonate solution to give a neutral aqueous phase (pH 7). The layers were separated and the ether phase was extracted with two 5 ml. portions of water to each of which was added sufficient bicarbonate solution to producean aqueous phase of pH 7. Evaporation of the combined aqueous solutions at low temperature and pressure left the sodium salt of a,a-dimethylbenzylpenicillin, which was finally dried in, a vacuum desiccator, yield 9.5 g.

The product inhibited Staph. Oxford at a concentration of 0.025 mcg./ml., the benzylpenicillin-resistant Staph. 1. at 25 mcg./ml., and the benzylpenicillin resistant Staph. 2. at 25 meg/ml.

Example 18.a,a-Diethylbenzylpenicillin 6-aminopenicillanic acid (5.6 g.) was acylated with a,a-diethylphenylacety1 chloride (6.0 g.) as described in Example 17 to give the sodium salt of u,a-diethylbenzylpenicillin (8.3 g.). i

It inhibited Staph. Oxford at a concentration of 0.05

mcg./ml., the benzylpenicillin resistant Staph. 1. at 2.5

rncg./ml., .and the benzylpcnicillin-resistant Staph. 2. at

Example 19. -a,a-Di-n-pr0pylbenzyIpen'icillin G-aminopenicillanic acid (7.55 g.) was acylated with a,a-di-n-propylphenylacetyl chloride (8.35 g.) as described in Example 17,-to give the sodiur n'salt of a,a-di-n-p ropylbenzylpenicillin (13.0 g.). T

It inhibited Staph.v Oxford at a concentration of 0.05 meg/ml, Staph. 1. at 1.25 meg/ml. and Staph. 2. at 1125 meg/ml.

Example 20.a,a-Di-n-bulylbenzylpenicillin 6-aminopenicillanic acid (3.9 g.) was acylated with u',a- ,;di-n-butylph enylacetyl chloride (4.8 g.) as described in Example 17 to give the sodium salt of a,a-di-n-butyibenzylpenicillin (6.5 g.).. a

It inhibited Staph. Oxford at 0.5 mcg./ml., Staph. 1. at 2.5 mcg;/ml., and Staph. 2. at 2.5 meg/ml.

Example 21.oe,u-Dicyclohexylbenzylpenicillin ,and. discarded, and the chloroform solution-Was washed with water (60 ml.). Sufiicient 8% aqueous sodium bicarbonatesolution was added to the chloroform solution to give, after vigorous shaking, a neutral emulsion (pH 7). This was evaporated at low temperature and pressure and the residual, white solid dried in a, vacuum desiccator.

.There was obtained 3.57 g. of the sodium salt of of Owl-di- 'cyclohexylbenzylpenicillin.

It inhibited Staph. Oxford at a concentration of 125 meg/ml, Staph. 1. at 62.5 mcg./ml., and Staph. 2. at 50 meg/ml. I I

a,a-Dicyclohexylphenylacetyl chloride, M.P. 9596 C.', was prepared by heating the corresponding acid in benzene with thionyl chloride and atrace of pyridine.

I H Example 22 1 In the procedure of Exampjle 17 the a,a-dimethy lphen- 1s ylacetyl chloride is replaced by an equirnolar weight of the acid chloride of 1a (4-nitrophenyl) -u-methylpropionic acid, u-(4- bromophenyl)-a-ethylpropionic acid, a-(4-t-butylphenyl) -a-isohexyl-n-butyr ic acid, a-(4-trifluoromethy1phenyl)-amethyl-isova1eric acid, a-(3-aminopheny-l)-a-methyl-n decanoic acid, a-(4-sulfamylphenyl)-a-methyl-a-cyclobutylacetic acid, a-(Z-benzylphenyl) -a-ethyl-a-cyclopentaylacetic acid, a- 3-methoxyphenyl -a-isopropyl-a-cyclohexylacetic acid, a-(2-iodophenyl)-a-n-butyl-u-cycloheptylacetic acid, a-(3-diethylaminophenyl) -a-methyl-n-butyric acid, a- (4-cyclohexylphenyl) -a-n-pr opyl-n-valeric acid, a-(4-methylaminophenyl)emu-dicychohexylacetic acid, 'a-(4-chlorophenyl') -a,a dicyclopentylacetic acid, a-(Z-acetamidophenyl) a-ethylpropionic acid, a-(3,4-dimethylphenyl)-u-ethyl-n-butyric acid, a-(3,4-dichlorophenyl)-a-ethyl-n=valcric acid, a-(3,4,5'-trimethoxyphenyl)-u-ethyl-n-hexanoic acid, a-(4-hydroxypheny1)-a-ethyl-n-octanoic acid, a(3,4-dihydroxyphenyl)-a-methyl-n-valeric acid, u-(4-methylsulfonylphenyl)w nghexpropionic acid and a-(2,5-dimethyl-4-hydroxyphenyl)-a-methylpropionic acid, 'respcctively,=to form the acids 6- [0(- (4,-nitrophenyl) a-methylpropionamido] penicil1anic acid, 6- oZ-( 4-bror'nophenyl -a-ethylpropionamido] penicillanic acid, 6-[a-(4t-butylphenyl)-a-isohexyl-n-butyramido]penicillanic acid, 6- a- (trifluoromethylphenyl -a-methyl-isovaleramido] penicillanic acid, 1 1 6-[a-(3-aminophenyl)-a-methyl-n-decanoamido]penicillanic acid, j 6- [oz- (4-sulfamylpheny1) -a-methyl-a-cyclobutylacetamido1penicillanic acid, 6-[a-(2-tbeniylphenyl)-a-ethyl-a-cyclopentacetamido] penicillanic acid; r I 6- [on-(3-methoxy'pheny1)-aisopropyl-a cyolohexylacet-' amido]penicillanic acid, 6-[a-(2 iodopheny)-ix-n-butyl-a-cycloheptylacetamido] penicillanic acid, 6-[w-(3-diethylaminophenyl)-a-methyl-n-butyramido] penicillanic acid, 6-[a-(4- cyclohexylphenyl)-a-n-propyl-n-valeramido] penicillanic acid,. 6- [a (4-methylaminophenyl) -a,a-dicyclohexylacetamido] penicillanic acid, 6-[a-(4-chlorophenyl)-a,a-dicyclopentylacetamido] penicillanic acid, 7 6- a- Z-acetamidophenyl) -a-ethylpropionamido] peni-v cillanic acid, 6- [a- 3 ,4- dimethylphenyl) -u-ethyl-n-butyramido penicillanic acid, r 6- [oc-( 3,4-dichlorophenyl) -'a-ethy1-n-valeramido] penicillanic acid, jv 6-[a-(3,4,5-trimethoxyphenyl)-a-ethyl-n-hexanoamido] penicillanic acid, 6-[a-(4 hydroxyphenyl)-a-ethyLn-octanoamido]penicillanic acid, I V 6- [cz- 3,4-dihydroxyphenyl) -a-methyl-n-valeramido] penicillanic acid, I 6- [oz- (4-methylthiophenyl) -a-methylpropionamido] penicillanic acid, 7, g 6- a- (4-methylsulfonylphenyl) -a-n-hexylpropionamido] penicillanic acid, and v, 6- [a- 2,5 -dimethyl-4-hydroxyphenyl) -u-methylpropionamido1penicillanic acid, respectively, which are isolated as their solid, water-soluble sodium salts, andsfound to inhibit Staph. aareus Smith as concentrations of 0.001 percent.

(d) TRIALKYL TYPE Example 23 Tert. butylpenicillin 6-aminopenicillanic acid (4.0 g.)- wastreated with trimethylacetyl chloride (2.5 vg.) as described in Example 1 to give the sodium salt of tertiary butyl penicillin as a yellow hygroscopic solid (3.7 g.). It inhibited Staph. aureus at a concentration of 2.5 mcg./ ml.

Example 24.Triethy'lmcthylpeiticillin A solution of triethylacetylchloride (0.98 g.) in dry chloroform ml.) was added dropwise during 5 mins. to a stirred mixture of 6-aminopenicillanic acid (1.3 g.), dry chloroform (30 ml.), and triethylamine (2.5 ml.). The mixture was stirred at room temperature for 2 /2 hrs. and then filtered. The filtrate was washed with 0.2 N hydrochloric acid (60 ml.) and then with water (60 ml.). Suflicient 3% aqueous sodium bicarbonate solution was added to the chloroform solution to give, after vigorous shaking, a neutral emulsion (pH 7). This was evaporated at low temperature and pressure and the residue was dried in a vacuum desiccator to leave the sodium salt of triethylmethylpenicillin as a white solid (1.84 g.).

It inhibited Staph. Oxford at a concentration of 0.05 mcg./ml., the benzylpenicillin-resistant Staph. 1. at 12.5 mcg./ml., and the benzylpenicillin-resistant Staph. 2. at 12.5 mcg./ml.

Example 25.Tri-n-pr0pylmethylpenicillin 6-aminopenicillanic acid (3.89 g.) Was acylated with tripropylacetyl chloride (3.68 g.) according to the method of Example 24 to give the sodium salt of tri-n-propylmethylpenicillin (5.98 g.).

It inhibited Staph. Oxford at a concentration of 0.05 mcg./ml., the benzylpenicillin-resistant Stap. 1. at 1.25 mcg./ml., and the benzylpenicillin-resistant Staph. 2. at 0.6 meg/ml.

Example 26.Tri-n-butylmethylpenicillin 6-aminopenicillanic acid (432 mg.) was acylated with tri-n-butylacetylchloride (493 mg.) according to the method of Example 24 to give the sodium salt of tri-nbutylmethylpenicillin as a colorless deliquescent solid (628 mg).

It inhibited Staph. aureus at a concentration of 0.6 mcg./ml., Staph. 1. at 2.5 mcg./ml., and Staph. 2. at 2.5 meg/ml.

Example 27 In the procedure of Example 24 the triethylacetyl chloride is replaced by an equimolar weight of the acid chloride of a,a-dimethyl-a-cyclohexylacetic acid, a-methyl-a-ethyl-a-cycloheptylacetic acid, a-n-propyl-a,ix-dicyclohexylacetic acid, a,o-di-n-propyl-a-cyclopentylacetic acid, a,a-di-n-hexyl-a-cyclobutylacetic acid u-n-hexyl-a-n-decyl-n-octanoic acid, 0:,oc-dilSO'dII1Yl-lSOVfil6ilC acid, a x,a-tricyclohexylacetic acid, a-methyl-a-n-propyl-n-decanoic acid, a,a-di-isopropyl-u-cyclohexylacetic acid, a,a-di-n-butyl-a-cycloheptylacetic acid, a,a-n-decyl-n-octanoic acid and a-methyl-a,a-dicyclohexylacetic acid, respectively, to form the acids 6-[a,a-dimethyl-a-cyclohexylacetamido]penicillanic acid, 6- a-methyl-a-ethyl-a-cycloheptylacetamido] penicillanic acid, 6-[can-propyl-a,a-dicyclohexylacctamido]penicillanic acid, 6- a,a-di-n-propyl-a-cyclopentylacetamido] penicillanic acid, 6-[a,a-di-n-hexyl-a-cyclobutylacetarnido]penicillanic acid, 6-[a-n-hexyl-a-n-decyl-n-octanoamido] penicillanic acid, 6-[a,a-di-isoamyl-isovaleramido] penicillanic acid, 6-[a,a,a-tricyclohexylacetamido] penicillanic acid, 6-[e-methyl-a-n-propyl-n-decanoamido]penicillanic acid,

i8 6-[a,a-di-isopropyl-a cyclohexylacetamido]penicillanic acid, V 6- a,a-di-n-butyl-a-cycloheptylacetamido] penicillanic acid, 6-[a,a-n-decyl-n-octanoamido]penicillanic acid, and 6- [a-methyl-a,a dicyclohexylacetamido] penicillanic acid, respectively, which are isolated as their solid, water-solu ble sodium salts and found to inhibit Staph. aureus Smith at concentrations of 0.001 percent.

(e) DIARYL (SUBSTTTUTED OXY) TYPE Example 28.a-Methoxydiphenylmethylpenicillin A solution-of a-methoxydiphenylacetyl chloride (1.04 g.) in dry chloroform (5 ml.) was added dropwise to a stirred mixture of 6-am'inopenicillanic acid (0.43 g.), chloroform (12 ml.), and 'triethylamine (0.84 ml.). Stirring was continued for 2 hours and the mixture was then filtered from a little undissolved solid. The filtrate was treated with successive small portions of 0.2 N hydrochloric acid, with vigorous shaking between additions, until the aqueous phase was at pH 23. The acid layer was separated and discarded, and the chloroform solution was washed with water (20 ml.). Enough 3% aqueous sodium bicarbonate solution was added to the chloroform solution to give, after vigorous shaking a neutral emulsion (pH 7). This -was evaporated at low temperature and pressure and the residual pale yellow solid was dried in a vacuum desiccator. There was obtained 0.90 g. of the sodium salt of u-methoxydiphenylmethylpenicillin.

The product inhibited Staph. aureus at a concentration of 0.01 mcg./ml., the benzylpenicillin-resistant Staph. 1. at 5 mcg./ml., and the benzylpenicillin-resistant Staph. 2. at 5 meg/ml.

The a-methoxydiphenylacetylchloride was prepared as a crude 'oil by warming a-methoxydiphenylacetic acid with thionyl chloride in benzene, followed by removal of solvent and exc'ess'r'eagent invacuo.

Example 29.--a.-lEtli0xydiphenylmethylpenicillin 6-aminopenicillanic acid (2.6 g.) was acylated with a-ethoxydiphenylacetyl chloride (3.3 g.) as described in Example 28 to give the sodium salt of wethoxydiphenyh methylpenicillin (2.8 g.).

It inhibited Staph. aa'reus at a concentration of 2.5 mcg./ml., Staph. 1. at 12.5 mcg./ml., and Staph. 2. at 12.5 meg/ml.

The a-ethoxydiphenylacetyl chloride was prepared as a crude oil by warming a-ethoxydiphenylacetic acid with thionyl chloride in benzene, followed by removal of solvent and excess reagent in vacuo.

Example 30.a-Is0prop0xydiphenylmethylpenicillin A solution of a-isopropoxydiphenylacetyl chloride (4.6 g.) in dry acetone (20 ml.) was added dropwise to a stirred solution of 6-aminopenicillanic acid (3.5 g.) and sodium bicarbonate (4 g.) in water (48 ml.) and acetone (28 ml.). When addition was complete the mixture was stirred at room temperature for 2 hours, then diluted with water (60 ml.) and extracted with ether ml. in 3 portions), only the aqueous phase being retained. This aqueous solution was covered with ether (30 ml.) and adjusted to pH 2 by the addition of N hydrochloride acid. After separating the layers, the aqueous phase was extracted with two 15 ml. portions of 'ether. The combined ether solutions (which at this stage contained the free penicillin acid) were Washed with water (50 ml.) and then shaken with sulficient 8% sodium bicarbonate solution to give a neutral aqueous phase (pH 7). The layers were separated and the ether phase was extracted with two 5 m1. portions of water to each of which was added enough bicarbonate solution to produce an aqueous phase of pH 7. The combined aqueous solutions were washed with ether (15 ml.) and evaporated at low temperature and pressure to leave the sodium salt of a-isopropoxydiphenylmethylpenicillin as a white solid, 'which was finally dried in a vacuum desiccator. Yield 2.66 g.

It inhibited Staph. Oxford at a concentration of 0.6 mcg./ml., Staph. 1. at 12.5 mcg./rnl., and Staph. 2. at 12.5 meg/ml.

The a-isopropoxydiphenylacetyl chloride was prepared as a crude oil by warming a-isopropoxydiphenylacetic acid with thionyl chloride in benzene, followed by removal of solvent and excess reagent in vacuo.

Example 31.a-Buloxydiphertylmethylpenicillin 6-aminopenicillanic acid (0.43 g.) was acylate-d with wbutoxydiphenylacetyl chloride (1 g.) as described in Example 28 to give the sodium salt of a-butoxydiphenylmethylpenicillin (1.2 g.).

It inhibited Staph. aureus at a concentration of 1.25 rncg./1nl., Staph. 1. at 2.5 meg/ml, and Staph. 2. at 2.5 meg/ml.

The wbutoxydiphenylacetyl chloride was prepared as a crude oil by warming oc-butoxydiphenylacetic acid with thionyl chloride in benzene, followed by removal of solvent and excess reagent in vacuo.

Example 32 In the procedure of Example 28 the ot-methoxydiphenylacetyl chloride is replaced by an equimolar weight of the acid chloride of a- 4-nitropheny1) -a-phenyl-a-cyclohexyloxyacetic acid,

a-(4-bIOIXlOPhE1'lYl o- (4-hydroxyphenyl) -a-cyclopentoxyacetic acid,

oc- (4-t-butylphenyl) -oc- (4-methoxyp'henyl) -u-cyclobutoxyacetic acid,

a- (4-tri-fluorornethylphenyl) -a- (4-methylphenyl) -amethoxyacetic acid,

- 3-aminophenyl) oc- (Z-hydroxyphenyl) -a-cyc1oheptoxyacetic acid,

ot- (4-sulfamylphenyl) -oc (Z-methoxyphenyl) -ocethoxyacetic acid,

a- (Z-benzylphenyl) -a- (Z-methylphenyl) -a-n-propoxyacetic acid,

u- 3 -methoxyphenyl) -a-p'henyl-a-n-hexoxyacetic acid,

a- 2-iodophenyl -0c- (3 -hydroxyphenyl -o-n-decoxyacetic acid,

(1(3 -diethylaminopheny-l -oc S-methoxyphenyl -ocisopropoxyacetic acid,

oc- (4-cyclohexylphenyl) -u- 3-methylphenyl) -a-nhexoxyacetic acid,

a,oL-di (4-met hylaminophenyl) -a-isoprop oxyacetic acid,

a- (4-chlorophenyl) -oc- (4-hydroxyphenyl) -a-methoxyacetic acid,

a-(2-acetarnidophenyl) -u- (3 -hydroxyphenyl) -x-ethoxyacetic acid,

a- (3 ,4-dimethylphenyl) -zx.- Z-hydroxyphenyl) -0L- methoxyacetic acid, v

a,ot-di- (3 ,4-dichlorophenyl) -a-cyclohexoxyacetic acid,

a,a-di- 3,4,5 -trirnethoxyphenyl) -a-cyclopentoxyacetic acid,

oc,oc-di- (4-hydroxyphenyl) -a-mcthoxyacetic acid,

a,Ct-dl- (3 ,4-di'hydroxypheny1)- x-ethoxyacetic acid,

04,0tdi- (4-methylthiophenyl) -u-1nethoxyacetic acid,

0.,oc-di- (4-methylsulfonylphenyl) -a-ethoxyacetic acid, and

-' 0t,a-(ll- (2,5 -dimethy1-4-hydroxyphenyl -u-methoxyacetic acid, respectively, to produce the acids 6- a- 4-nitrophenyl) -a-phenyl-a-cyclohexyloxyacetamido] penicillanic acid,

6- oc- 4-bromophenyl -a- 4-hydroxyphenyl) -u-cyclopentoxyacetamido]penicillanic acid,

6- a- (4-t-butylphenyl) -a-(4-methoxyphenyl -a-cyclobutoxyaceta-mido]penicillanic acid, 7

6- [a-(4-tri-fiuoromethylphenyl) -a-(4-methylpheny1) -oz.-

nlethoxyacetamido] penicillanic acid,

6- [u- 3-arninophenyl a- 2-hydroxyphenyl) -l2.-CyC10- heptoxyaeeamido] enicillanic acid,

6- a- (4-sulfamylphenyl -oc- ('2-methoxyphenyl -aethoxyacetamido] penicil-lanic acid,

6- u- (2-benzylphenyl) -oc- (Z-methylphenyl) -a-n-propoxyacetamido] penicillanic acid,

6- [a- 3-methoxyphenyl) -a-phenyl-a-n-hexoxyacetamido]penicillanic acid,

6- a- ('2-icdophenyl -a-( 3 -hydr-oxyp-henyl) -a-n-dec-oxyaceta'mido] penicillanic acid,

6- oc- 3 -die-thy1aminophenyl) -a- (3 -methoxyphenyl) -ocisopropoxyacetamido] penicillanic acid,

6- oc- 4-cyclohexylphenyl) -a- 3-rnethylphenyl) -a-nhexoxyacetamido]penicillanic acid,

6- [0a,oL-di- (4-rnethylaminophenyl -a-ispr opoxyacetamido]penicillanic acid,

6- (X- 4-chlorophenyl -OL- (4-hydroxyphenyl) -a-methoxyacetamido]penicillanic acid,

6- c4- Z-acetamidophenyl -oc- 3-hydroxypheny1) oc ethoxyacetamido] penicillanic acid,

6- [a- 3 ,4-dimethylphenyl -a (2-hydroxyphenyl) -ocmethoxyacetamido]penicil-lanic acid,

6- [a,a-di- (3 ,4-dichloropheny1) -a-cyclohexoxyacetamido]penicil1anic acid,

6-[a,a-di-(3,4,5-trimethoxyphenyl)-a-cyclopentoxyacetam-ido1penicillanic acid,

6- oc,oc-di- (4-hydroxyphenyl) -a-methoxyacetamido] penicillanic acid,

6-[c m-di-(3,4-dihydroxyphenyl)-u-ethoxyacetamido] penicillanic acid,

6- oc,ocdi (4-methylthiophenyl) -a-rnethoxyacetarnido] penicillani-c acid,

6- 0c,zx-di- (4-methylsulfonylphenyl) -a-ethoxyacetamido] pen-icillanic acid,

6- 0:,04-di- (2,5 -dimethyl-4-hydroxyphenyl) -a-met=hoxyacetamide]penicillanic acid, respectively,

which are isolated as their solid, water-soluble sodium salts and found to inhibit Staph. aureus Smith at concentrations of 0.001 percent by weight.

( DIARYL (SUBSTITUTED THIO) TYPE Example 33.ot'Ethylthiodiphenylmethylpenicillin A solution of a-ethylthiodiphenylacetyl chloride (8.0 g.) in dry acetone (30 ml.) was added dropwise during 10 .mins. to a stirred solution of 6-aminopencillanic acid (5.4 g.) and sodium bicarbonate (6.3 g.) in water ml.) and acetone (60 ml.). When addition was complete the mixture was stirred at room temperature for 2 /2 hrs., then diluted with water (50 ml.) and extracted with ether (2 x ml.), only the aqueous phase being retained. This aqueous solution was covered with ether (50 ml.) and adjusted to pH 2 by the addition of N hydrochloric acid. After separating the layers, the aqueous phase was extracted with two further 25 ml. portions of ether. The combined ether solutions (which at this stage contained the free penicillin acid) were washed with water (50 ml.) and then shaken with sufficient 8% sodium bicarbonate solution to give a neutral aqueous phase (pH 7). The layers were separated and the ether phase Was extracted with two 5 ml. portions of water to each of which was added sufiicient bicarbonate solution to produce on aqueous phase of pH 7. The combined aqueous extracts were Washed with ether (25 ml.) and then evaporated at low temperature and pressure to leave the sodium salt of a-ethylthi0diphenylmethylpenicillin as a pale yellow solid which was finally dried in a vacuum desiccator. Yield 8.67 g.

The product inhibited Staph. Oxford at a concentration of 0.025 rncg./rnl., the benzylpenicillin-resistant Staph. 1. at 1.25 mcg./.ml., and the benzylpenicillin-resistant Staph. 2. at 2.5 meg/ml.

The a-ethylthiodipihenylacetyl chloride Was prepared as a crude oil by warming the corresponding acid with thiony-l chloride in benzene, then removing the solvent and excess reagent in vacuo. oc-Ethylthiodiphenylacetic acid, MP. 131-132" C., was itself prepared by condensing Example 34.a-n-Prpylthz'odiplzenylmethylpenicillin G-aminopenicillanic acid (6.25 g.) was .acylated with a-n-propylthiodiphenylacetyl chloride (8.8 g.) according to the procedure described in Example 33 to give the sodium salt of a-n-propylthiodiphenylmethylpenicillin as a White powder (8.36 g.).

It inhibited Staph. Oxford at a concentration of 0.6 mcg./ml., Staph. 1. at 2.5 mcg./ml., and Staph. 2. at 1.25 mcg/ml.

The ct-n-propylthiodiphenylacetyl chloride was prepared as a crude oil by warming the corresponding acid with thionyl chloride in benzene, then removing the solvent and excess reagent in vacuo. ct-n-Propylthiodiphenylacetic acid, M.P. 116-118" C., was itself prepared by condensing benzilic acid with n-propylmercaptan according to the general method of Barkenbus and Brower (J. Amer. Chem. Soc., 77, 579, 1955).

Example 35. x-n-Bmylthi0dlphenylmeihylpenicillin A solution of a-n-butylthiodiphenylacety-l chloride (0.64 g.) in dry chloroform (6 ml.) was added dropwise to a stirred mixture of 6-aminopenicillanic acid (0.43 g.), chloroform (12 ml.), and triethylamine (0.84 ml.). The mixture was stirred at room temperature for 2 /2 hours and then filtered. The filtrate was treated with successive small portions of 0.2 N hydrochloric acid, with vigorous shaking between additions, until the aqueous phase was at pH 2. The acid layer was separated and discarded, and the chloroform solution was washed with water (15 ml.). Sufficient 3% aqueous sodium bicarbonate solution was added to the chloroform solution to give, after vigorous shaking, a neutral emulsion (pH 7). This was evaporated at low temperature and pressure and the residual white solid dried in a vacuum desiccator. There was obtained 0.62 g. of the sodium salt of a-n-butylthiodiphenylmethylpenicillin.

It inhibited Staph. aureus at a concentration of 1.25 mcg./ml., Staph. 1. at 2.5 meg/ml, and Staph. 2. at 2.5 meg/ml.

The a-n-butylthiodiphenylacetyl chloride was prepared as a crude oil by warming the corresponding acid, itself prepared by the method of Barkenbus and Brower (J. Ame-r. Chem. Soc., 77, 579, 1955), with thionyl chloride in benzene, then removing the solvent and excess reagent in vacuo.

Example 36.u-Is0pr0pyltlziodiplzenylmethylpenicillin 6-aminopenieillanic acid (6.5 g.) was acylated with ot-isopropylthiodiphenylacetyl chloride (10.4 g.) according to the procedure described in Example 33 to give the sodium salt of ot-isopropylthiodiphenylmethylpenicillin as a White solid (9.65 g.).

It inhibited Staph. Oxford at a concentration of 0.6 meg/ml, Staph. 1. at 5 mcg./ml., and Staph. 2. at 5 meg/ml.

The a-isopropylthiodiphenylacetyl chloride was prepared as a crude oil by Warming the corresponding acid with thionyl chloride in benzene, then removing the solvent and excess reagent in vacuo. u-Isopropylthiodiphenylacetic acid, M.P. 130132 C., was itself prepared by condensing benzilic acid with isopropyl mercaptan ac cording to the general method of Barkenbus and Brower (J. Amer. Chem. Soc., 77, 579, 1955).

Example 37.-oc-S -buzyltlziodiplz-enylmetlzy[penicillin 6-aminopenici1lanic acid (5.4 g.) was acylated with asec-butylthiodiphenylacetyl chloride (9.6 g.) according to the procedure described in Example 33 to give the sodium salt of a-sec-butylthiodiphenyimethylpenicillin as a 'pale yellow solid (7.87 g).

It inhibited Staph. Oxford at a concentration of 0.6

22 mcg./ml., Staph. 1. at 5 mcg./ml., and Staph. 2. at 5 meg/ml.

The u-sec-butylthiodiphenylacetyl chloride was prepared as a crude oil by warming the corresponding acid with thionyl chloride in benzene, then removing the solvent and excess reagent in vacuo. e-sec-butylthiodiphenylacetic acid, M.P. 97 C., was itself prepared by condensing benzilic acid with sec-buty-lmercaptan according to the general method of Barkenbus and Brower (J. Amer. Chem. Soc., 77, 1955, 579).

Example 38.-a-Benzyltlziodiphenylmetlzylpenicillin 6-aminopenicillanic acid (4.95 g.) was acylated with abenzylthiodiphenylacetyl chloride (8.85 g.) according to the procedure described in Example 33 to give the sodium salt of a-benzylthiodiphenylmethylpenicillin (7.53 g.).

It inhibited Staph. Oxford at a concentration of 0.125 mcg./ml., Staph. 1. at 0.5 meg/ml, and Staph. 2. at 0.6 meg/ml.

The otbenzylthiodiphenylacetyl chloride was prepared as a crude oil by warming the corresponding acid, itself prepared by the method of Barkenbus and Brower (J. Amer. Chem. Soc., 1955, 77, 579), with thionyl chloride in benzene, then removing the solvent and excess reagent in vacuo.

Example 39 In the procedure of Example 33 the a-ethylthiodiphenylacetyl chloride is replaced by an equimolar weight of the acid chloride of a- (4nitrophenyl -a-phenyl-a-cyclohexylthioacetic acid,

66- 4-bromophenyl) -a- (4-hydroxypheny1) -a-cyclopentylthioace'tic acid,

oz- (4-t-butylphenyl) -a- 4-methoxyphenyl -oz-cyclobutylthioacetic acid,

06- (4-trifluoromethylphenyl) -u- (4-methylphenyl) -oc methylthioacetic acid,

ot-(3-aminophenyl) -a-(2-hydroxypheny1) -a-cycloheptylthioacetic acid,

OL- 4-sulfamylphenyl o- 2-methoxyphenyl) -a-ethylthioacetic acid,

a(2-benzylphenyl -u- Z-methylphenyI) -u-n-propylthioacetic acid,

a- (3 -rnethoxypheny1) -a-phenyl-ot-n-hexylthioacetic acid,

00- (2-iodophenyl) -0- 3-hydroxyphenyl) -u-n-decylthioacetic acid,

ot-(3-diethylaminophenyl)-1x-(3-metl1oxyphenyl) -a-iSO- propylthioacetic acid,

OL-(4-CYClOh6XYlPh6l1Yl) -:x- 3-methylphenyl) -a-n-hexylthioacetic acid,

a,o-di-(4-methylaminophenyl)-ot-isopropylthioacetic acid,

a- (4-chlorophenyl) x- (4-hydroxyphenyl -a-rnethylthioacetic acid,

a-(2-acetamidophenyl) -0L-(3-hYdIOXYPhEDYl)-0-etl1ylth O- acetic acid,

u-(3,4-dimethylphenyl)-a-(2-hydroxyphenyl)-a-methylthioacetic acid, 0L,0-dl-(3,4-dlChlOIOph6nyl)-0L-CYClOl'l6X- ylthioacetic acid,

east-di-(3,4,5-trimethoxyphenyl)-a-cyclopentylthioacetic acid,

a,a-di-(4-'hydroxyphenyl)-a-methylthioacetic acid,

oc,oc-di 3,4-dihydroxyphenyl -ot-ethylthioacetic acid,

a,a-di- (4-methylthiophenyl) -a-methylthioacetic acid,

a,a-di- 4-methylsulfonylphenyl -a-ethylthioacetic acid,

and

a x-di-(2,5-dimethyl-4-hydroxyphenyl)-u-methylthioacetic acid, respectively to produce the acids 6- oc- 4-nitrophenyl flit-phenyl-a-cyclohexylthioacetamidoJpenicillanic acid,

6-[a-(4-b1'omophenyl) -ot-(4-hydroxyphenyl)-a-cyclopentylthioacetamido]penicillanic acid,

6- a- (4-t-butylphenyl) -0e (4-methoxyphenyl) -o-cyclobutylthioacetamido]penicillanic acid,

6- a-'( 4-tri-fiuoromethylphenyl -OL- (4-methylphenyl) -amethylthioacetamido]penicillanic acid,

25 6- [L,oc-di 4-hydroxyphenyl) -a-phenylthioacetamido] penicillanic acid, 6- (x,adi-( 3,4-dihyd'roxyphenyl -a-phenylthioacetamido] penicillanic acid, 6-[ x, z-di-(4-methylthiophenyl)'-a-phenylthioacetamido] penicillanic acid, 1 v 6-[a,oc-di-(4-methylsulfonylphenyl)-a-phenylthioace-' tamido]penicillanic acid and I v 6-[e codi-(2,5-dimethyl-4-hydroxyphenyl)-a-phenylthioacetamidoJpenicillanic acid, respectively, which are isolated as their solid, water-soluble sodium salts and found to inhibit Staph. aureus Smith at concentrations of 0.001 percent by weight.

(g) DIALKYL (SUBSTITUTED OXY) TYPE Example 43.-ot-Phenoxyisopropylpenicillin A solution of ix-phenoxyisobutyryl chloride (5.77 g.) in dry acetone (40 ml.) was added dropwise during mins. to a stirred solution of 6-aminopenicillanic acid (5.63 g.) and sodium bicarbonate (6.6 g.) in water (90 ml.) and acetone (50 ml.). When addition was complete the mixture was stirred at room temperature for 2 hours then diluted with water (50 ml.) and washed with two 90 ml. portions of ether. The aqueous phase was brought to pH 2 by addition of N hydrochloric (57 ml.) and the liberated penicillin-free acid was extracted into ether (3 x 25 ml.). These ether extracts were washed with water (2 x ml.) and then shaken with sufficient 8% sodium bicarbonate solution to give a neutral aqueous phase (pl-I 7). The layers were separated and the ether phase was extracted with two 5 ml. portions of water to each of which was added sulficient bicarbonate solution to produce an aqueous phase of pH 7. Evaporation of the combined aqueous solutions at low temperature and pressure left the sodium salt of 6-(or-phenoxy-a-methylpropionamido)penicillanic acid as a white solid, which was finally dried in a vacuum desiccator. Yield 10.6 g.

The product inhibited Staph. Oxford at 0.005 mcg./ml.

The a-phenoxyisobutyryl chloride was prepared from a-phenoxyisobutyric acid by the method of Gabriel, Ben, 46,1347 (1913).

Example 44 A solution of a-phenoxyisobutyric acid (0.72 g.) and triethylamine (0.55 ml.) in dry acetone (8 ml.) was stirred and cooled to 0 C. A solution of sec. butyl chlorocarbonate (0.51 ml.) in dry acetone (8 ml.) was added dropwise over 5 minutes after which the mixture was stirred at 0-5 C. for 1 hour. A solution of 6-aminopenicillanic acid (0.86 g.) in 0.5 N sodium bicarbonate solution (16 ml.) was added to the suspension, and the resulting clear solution was stirred at room temperature for 2 hours. It was washed then with ether (2 x 15 ml.) and the washings were discarded. The aqueous phase was then acidified to pH 2.5 and 6-(a-phenoxy-a-methylpropionamido)penicillanic acid isolated as in Example 43. T he yield of sodium salt was 1.29 g.

Example 45 Isobutyl chloroformate (0.1 mol.) was added to a chilled solution of a-phenoxy-isobutyric acid (0.1 mol.) and triethylamine (0.1 mol.) in 250 ml. of acetone and 500 ml. of dioxane and the resulting mixture was stirred for approximately thirty minutes at about 3 C. A chilled solution of 6-aminopenicillanic acid (0.1 mol.) and triethylamine (0.1 mol.) in 500 ml. of water was then added to the above acylating mixture and this reaction mixture was stirred for approximately one hour in the cold. After the addition of a saturated solution of sodium bicarbonate, the alkaline reaction mixture was extracted with ether and the ethereal extract was discarded. After covering the aqu ous solution with methyl isobutyl ketone, it was acidified to a pH of about 2 with 6 N hydrochloric acid and was then extracted twice with methyl isobutyl ketone. The combined extracts, in which the product 6-(d-phenoxy-amethylpropionamido)penicillanic acid was contained, were 26 washed with water, dried over anhydrous sodium sulphate and filtered. The addition of 40 ml. of a 40% butanolic solution of potassium Z-ethylhexauoate caused the precipitation of the potassium salt of the above penicillanic acidi After tr'ituration' with ether, this potassium salt was dried invacuo over phosphorus pentoxide, recovered as a water-soluble powder, andfound to weigh 20.8 grns. (0.05 mol.), to melt with decomposition at 232-234 C., to contain the B-lactar'n structure as shown by infrared analysis and to inhibit Staph. aureus Smith at a concentration of 0.4 mc'g./ml., and to exhibit versus Staph. am'eus Smith upon intramuscular injection in mice CD of 3.7 mgmJkg.

Example 46 w A- solution of v;-(2,4-dichlorophenoxy)-m-benzyl-nbutyryl chloride (10.3 g.) in dry acetone (40 ml.) was added dropwise during 15 minutes to a stirred solution of 6-aminopenicillanic acid (5.63 g.) and'sodium bicarbonate (6.6 g.) in water (901111.) and acetone (50 ml.). When addition was complete, the mixture was stirred at room temperature for 2 hours then diluted with water (50 ml.) and washed with two ml. portions of ether. The aqueous phase was brought to pH 2 by addition of N hydrochloric acid (57 ml.) and the liberated penicillin-free acid was extracted into ether (3 x 25 ml.). These ether extracts were washed with water (2 x 20 ml.) and then shaken with sufiicient 8% sodium bicarbonate solution to give a neutral aqueous phase (pH 7). The layers were separated and the ether phase was extracted with two 5 ml. portions of water to each of which was added sufiicient bicarbonate solution to produce an aqueous phase of pH 7. Evaporation of the combined aqueous solution at low temperature and pressure left the sodium salt of 6-[a-(2,4-dichlorophenoxy)-a-benzy1-n butyramido]penicillanic acid as a white solid, which was finally dried in a vacuum desiccator.

The product inhibited Staph. aureus Smith at a concentration of 0.001 percent by weight.

Example 47 Isobu-tyl chloroformate (0.1 mol.) was added to a cooled (03 C.) solution of a-(4-hydr'oxyphenoxy)-amethyl-isovaleric acid (0.1 mol.) and triethylamine (0.1 mol.) in 250 ml. of acetone and 500 ml. of dioxane and the resulting mixture was stirred for thirty minutes in the cold. This mixture was then treated with a cool solution of 6-aminopenicillanic acid (0.1 mol.) and triethylamine (0.1 mol.) in 500 ml. of water and was stirred in the cold for an additional hour. The pH of the reaction mixture was adjusted to approximately 8 by the addition of a saturated solution of sodium bicarbonate and was then extracted with ether. Following the ether extraction the aqueous solution was covered with methyl isobutyl ketone and was acidified to a pH of 2 by the addition of 6 N HOl. The acidified aqueous solution was then extracted twice with methyl isobutyl ketone. The combined extracts, in which the product 6-[a-(4-hydroxy phenoxy)-u-me-tl1yl-iso-valeramido]penicillanic acid was contained, were washed with water, dried over anhydrous sodium sulphate and filtered. The addition of 40 ml. of a 40% butanolic solution of potassium Z-ethylhexanoate caused the precipitation of the potassium salt of the above penicillanic acid. After trituration with ether, this potassium salt was dried in vacuo over phosphorous pentoxide and recovered as a water-soluble powder which inhibited the growth of Staph. aureus Smith at a concentration of 0.001 percent by weight.

Example 48 After the addition of isobutyl ohloroforimate (0.1 mol.) to a chilled (4 C.) solution of a-(2,4-dimethoxyphenoxy)-u-methyl-n-valer-ic acid (0.1 mol.) and triethylamine (0.1 mol.) in 250 ml. of acetone and 500 ml. of dioxane, the reaction mixture was stirred -for approximately thirty minutes in the cold. This reaction mixture was then treated with a cool (5 C.) solution of 6-aminopenicil- 'lanic acid (0.1 mol.) and triet-hylamine (0.1 nrol.) in 500 ml. of water and stirring was continued for an additional hour. Upon completion of the stir-ring period, the reaction mixture was made slightly basic (pH=8) by the addition of a saturated sodium bicarbonate solution and was then extracted with ether. The ethereal extract was discarded, the aqueous alkaline solution was covered with methyl isobutyl ket-one and was then acidified (pl-i=2) with 6 N hydrochloric acid. The acidified aqueous solution was extracted twice with methyl isobutyl ketone and these extracts were combined. These combined extracts were washed with water, dried over anhydrous sodium sulphate and filtered. The 6-[a(2,4-dlinethoxyphenoxy)-u-methyl n valeramido1-penici1lanic acid was precipitated in the form of its potassium salt, by the addition of 40 ml. of a 40% butanolic solution of potassium Z-ethylhexanoate. A-fter tr-ituration with ether, this potassium salt was dried in vacuo over P and recovered as a water-soluble powder which inhibited the growth of Staph. aureus Smith at a concentration of 0.001 percent by weight.

Example 49 A solution of a-phenoxy-aabenzylphenylacetyl chloride (9.7 g.) in dry acetone (40 ml.) was added dropwise during 15 minutes to a stir-red solution of 6-aminopenicillanic acid (5.63 g.) and sodium bicarbonate (6.6 g.) in water (90 ml.) and acetone (50 ml.). When addition was complete the mixture was stirred at room temperature for 2 hours then diluted with water (50 ml.) and washed with two 90 ml. portions of ether. The aqueous phase was brought to pH 2 by addition of N hydrochloric acid (57 ml.) and the liberated penicillin-free acid was extracted into other (3 x 25 ml.). These ether extracts were washed with water (2 x 20 ml.) and then shaken with suflicient 8% sodium bicarbonate solution to give a neutral aqueous phase (pH 7). The layers were separated and the ether phase was extracted with two 5 ml. portions of water to each of which was added suflicient bicarbonate solution to produce an aqueous phase of pH 7. Evaporation of the combined aqueous solutions at low temperature and pressure left the sodium salt of 6-(aphenoxy-wbenzylphenyl-ace-tamido)penicillan-i-c acid as a white solid which was finally dried in a vacuum desiccator.

The product inhibited Staph. aureus Smith at a concentration of 0.001 percent by weight.

Example In the procedure of Examples 43 to 45 the acid was replaced by a-phenoxy-ot-benzylpropionic acid, or-phenoxy-a-benzylphenylbutyric acid, ot-(4-nitrophenoxy)-e-methyl-n-butyric acid, a-(4-bromophenoxy)-aphenyl-n-valeric acid, a-(4-tertiary butylphenoxy)-a benzyl-n-butyric acid, or-(4-trifluoromethylphenoxy)- x-phenyl-n-butyric acid, a-(3-aminophenoxy)-a-benzy1-n-valeric acid, a-(4-sulphamylphenoxy)-ot-methylpropionic acid, u-(2-benzylphenoxy) -a-methyl-n-butyr-ic acid, a-(3-methoxyphenoxy) -a-methyldecanoic acid,

11- (Z-iodophenoxy) haphenylpropionic acid, a-(3-diethylaminophenoxy) -a-methyl-n-butyric acid, e-(4-cyclohexylphenoxy)-a,u-dimethylacetic acid, ct-(3-aminophenoxy)- x-methylpropionic acid, a-(2,3,4,5,6-pentachl-orophen-oxy)isobutyric acid, and a-(2,4-diisoa1nylphenoxy) -m-methyl-n-butyric acid,

respectively, to produce the acids' 6-(a-phenoxy-a-benzylpropionamido)penicillanic acid,

6- a-phenoxy-a-benzylphenylbutyramido penicill anic acid, l

6- a- (4-nitro-phenoxy -ot-me'thyl-n-butyramido] penicillanic acid, i

6- [oc- (4-bromophenoxy) -ot-phenyl-n-valeramido] penicillanic acid,

iii

6- [a- 4-tertiary-butylphenoxy) -a-benzyl-n-butyramido] penicillanic acid,

6- [.1- (4-trifiuoromethylphenoxy -u-phenyl-n-butyramidolpenicillan-ic acid,

6- u- 3-amin0phenoxy) -a-benzyl-n-vale-ramido] penicillanic acid,

6- a- (4-sulphamylphenoxy) -a-methylpropionamido] penicillanic acid,

6- [oc- Z benzylphenoxy) -u-methyl-n-butyramido] penicillanic acid,

6- OL- 3-methoxyphenoxy) -u-methyldecanoam-ido] penicillanic acid,

6- [oc- (2-iodophenoxy -a-phenylpropionamido] penicillanic acid,

6- u- 3-diethylaminophenoxy) -a-methyl-n-butyramido] penicillanic acid,

6- a- 3 -cyclohexyl-phenoxy)-a,a-di1nethylacetamido]- penicillanic acid,

6- a- 3 -aminophenoxy) -a-methylpropionamido] penicillanic acid,

6- oc-Z, 3 ,4,5,6-pentachlorophenoxy) isobutyramido] penicillanic acid and 6- oc- (2,4-di-isoamylphenoxy -a-methyl-n-butyrarnido] penicillanic, acid,

respectively, which were isolated as their solid,-watersoluble potassium salts and found to inhibit Staph.

aureus Smith at a concentration of 0.001 percent by weight.

([1) DIALKYL (SUBSTITUTED THIO) TYPE Example 51 .a-Phenylthioisopropylpenicillin A solution of a-phenylthioisobutyryl chloride (6.14 g.) in dry acetone (30 ml.) was added dropwise during 10 minutes to a stirred solution of 6-arninopenicillanic acid (5.63 g.) and sodium bicarbonate (6.55 g.) in water ml.) and acetone (60 ml.). When addition was complete the matter was stirred at room temperature for 2 hrs., then extracted with ether (2 x 90 ml.), only the aqueous phase being retained. This aqueous solution was covered with ether (50 ml.) and adjusted to pH 2 by the addition of N hydrochloric acid. After separating the layers, the aqueous phase Was extracted with two further 25 ml. portions of ether. The combined ether solutions (which at this stage contained the free penicillin acid) were washed with water (50 ml.) and then shaken with suflicient 8% sodium bicarbonate solution to give a neutral aqueous phase (pH 7). The layers were separated and the ether phase was extracted with two 5 ml. portions of water to each of which was added sufficient bicarbonate solution to produce an aqueous phase of pH 7. The combined aqueous extracts were washed with ether (30 ml.) and then evaporated at low temperature and pressure. The residue was finally dried in a vacuum desiccator to give the sodium salt of oc-phenylthioisopropylpenicillin as a White solid (8.7 g.).

The product inhibited Staph. Oxford at a concentration of 0.012 meg/ml, and the benzylpenicillin-resistant Staph. 1 at 25 mcg./ml., and the benzylpenicillin-resistant Staph. 2 at 12.5 mcg./ml.

Example 52.-a-Ethyl-ot-phenylzhiopropylpenicillin 6-aminopenicillanic acid (6.49 g.) was acylated with ot-ethyl-a-phenylthiobutyryl chloride (7.28 g.) according to the procedure of Example 51 to give the sodium salt of a-ethyl-a-phenylthiopropylpenicillin as a butt-colored solid (12.7 g.).

It inhibited Staph. Oxford at a concentration of 0.05 mcg./ml., Staph. 1 at 5 meg/ml, and Staph. 2 at 2.5 mcg./ml.

a-Ethyl-oz-phenylthiobutyryl chloride, B.P. 86 C./0.05 mm., was prepared from the corresponding acid and thionyl chloride. a-Ethyl-os-phenylthiobutyric-acid, M.P. 7374 C., was itself obtained by the action of thiophenol on ethyl e-bromo-a-ethylbutyrate in the presence of aqueous alcoholic sodium hydroxide.

Example 53.u-Prpylx-phenylthiobutylpenicillin 6-amin0penicillanic acid (4.32 g.) Was acylated with a-propyl-a-phenylthiovalery1 chloride (5.41 g.) according to the procedure of Example 51 to give the sodium salt of a-propyl-o-phenylthiobutylpenicillin (8.6 g.).

It inhibited Staph. Oxford at a concentration of 0.5 rncg./'ml., Staph. 1 at 5.0 mcg./ml., and Staph. 2 at 1.25 rncg./ ml. I

a-Propyl-u-phenylthiovaleryl chloride, B.P. 120 C./ 0.1 mm., was prepared from the corresponding acid and thionyl chloride. a-Propyl-a-phenylthiovaleric acid, M.P. 118 C., was itself obtained by the action of thiophenol on ethyl a-bromo-oc-propylvalerate in the presence of aqueous alcoholic sodium hydroxide.

Example 54 In the procedure of Example 51 the a-phenylthioisobutyryl chloride is replaced by an equimolar weight of the acid chloride of u-(4-nitrophenylthio)-ot-methylpropionic acid,

a- 4-b'r-omophenylthio -a-ethylpropi'onic acid,

a- (4-t4butylphenylthio a-isohexyl-n-butyric acid,

a-(4-trifluoromethylphenylthio)-u-methyl-isovaleric acid,

a- 3-aminophenylthio -a-rnethyl-n-decanoic acid,

a- 4-sulfamylphenylthio) -aY-methyLor-cyclobutylacetic acid,

oz- (2-benzylphe'nylthio) -a-ethy1-a-cyclopentylacetic acid,

a- 3-methoxyphenylthio -a-isopropyl-ci-cyclohexylacetic E-(Z-iodophenylthio)-a-n-butyl-u-cycloheptylacetic acid,

a-(3-die'thylam'inophenylthio) -a-methyl-n-butyric acid,

a-(4-cyclohexylphenylthio)-a-n--propyl-n-valeric acid,

u-(4-methylaminophenylthio)-a,ot-dicyclohexylacetic acid,

a-(4-chlorophenylthio)-a,a-dicyclopentylacetic acid,

a-(Z-acetamidophenylthio -a-ethy1propionic acid.

a- 3,4-dimethy'lphenylthio -u-ethyl-n-butyric acid,

a-( 3,4dichlorophenylthio') -a-ethyl-n-valeric acid,

11- 3,4,5 -trimethoxy phenylthio -a-ethyl-n-hexanoic acid,

a-(4-hydroxyphenylthio)-a-ethyl-n-octanoic acid,

a- 3,4 dihydroxyp henylthio -a-'methyl-n-valeric acid,

a-(4-methy1thiophenylt'hio)-u-methylpropionic, acid,

u-(4-rnethy1sulfonylphenylthio)- x-n-hexylpropionic acid and 06- (2,5 -dimethyl-4-hydroxyphenylthio) -u-m'ethyl propionic acid,

respectively, to form the acids 6- oc- (4-n-itrophenylthio -u-methylpropio'namido 1 penicillanic acid,

6- a- 4-b-rornophenylthio) -a-ethylpropionamido] -peni cillanic acid,

6- a- (4-t-butyl-phenylth-io) wis'oheXyl-n-butyramido penicilla'ru'c acid,

6- atrifiuoromethylphenylthio) -a-methylisovalera'mid01penioillanic acid,

6- [a-( S-aminophenylthio) -u-methyl-n-decanoamido] penicillanic acid,

6- [a- (4-sulfamylphenyl -a-'methyl-a-cyclobuty-lacetamido]penicillanic acid,

6-[a-(2-benzylphenylthio)-a-ethyl-a-cyclopentylacetamidolpenicillanic acid,

6-[a-(3-methoxyphenylthio)-a-isopropyl-ot-cyclohexylacetamido]penicillanic acid,

6- oz- (2-idodophenylthio) -a-n-butyl-a-cycloheptylacetamido]penicillanic acid,

6- a- 3-diethylaminophenylthio) -o-methyl-nbutyrarnid'o] penicillanic acid,

6- a- (4-cyclohexylp-henylthio) -u-n-propyl-n-'valeramidoJpenicillanic acid,

6- a- (4-methyl-aminophenylthio) -a,a-dicyclohexylacetarn'ido1penicillanic acid,

6- a- (4-chlorophenylthio) -a,a,-dicyclopentylacetamido] penioillanic acid,

6-[u-(2-acetamidophenylthio)-a-ethylpropionamido]- penicillanic acid,

30 6- [oc- 3,4-dimethylphenylthio) aa-ethyl-n-butynamgido] penicillanic acid, 6- oc- 3 ,4-diohlorophenylthio) -a-ethyl-n-'v-aleramido] penicillanic acid, 6- u- 3,4,5 -trimethyloxyphenylthio) methyl-nhexanoamid0]penicillanic acid, 6- or (4 -hydroxyph'enylthio) -a-ethyl-n-'o'ctanoamido]penioillanic acid, 6- [cc-( 3,4-dihydroxyphenylth-io) -a-methyl-n-valeramido] penicillanic acid, 6- a- 4-me thylthiophenyl-thio) -a-methylpropionamido] penicillanic acid, 6- Ol- (4-methylsulfonylpheuylthio -a-n-hexylpropionamido]penicillanic acid, and 6- [a- 2,5 -dimethyl-4-hydroxy-phenylthio) -u-m'ethylprop-ionamido]penicillanic acid, respectively, which are isolated as their solid, water-soluble sodium salts and found to inhibit Staph. aureus Smith at concentrations of 0.001 percent.

(j) DIARL HETERO'CYCDLIC TYPE Example ;a,a-DiphenyI-Z-thienyImethylpenicillin A solution of et-2-thienyldiphenylacetyl chloride (6.3 g.) in dry acetone (40 ml.) was slowly added to a stirred solution of 6-aminopenicillanic acid (4.33 g.) and sodium bicarbonate (5 g.) in water (100 ml.) and acetone ml.). When addition was complete the mixture was stirred at room temperature for 3 hours, and then filtered. The filtrate was extracted with ether (2 x 100 ml.) and the extracts were discarded. The aqueous phase was covered with ether (50 ml.) and adjusted to pH 2 by addition of N hydrochloric acid. After separating the layers, the aqueous phase was extracted with two more 25 ml. portions of ether. The combined ether solutions (which at this stage contained the free penicillin acid) were washed with water (50 ml.) and then shaken with suificient 8% sodium bicarbonate solution to give a neutral aqueous phase (pH 7). The layers were separated and the ether phase was extracted with two 5 m1. portions of water to each of which was added enough bicarbonate solution to produce an aqueous phase of pH 7. The combined aqueous extracts were washed with ether, then evaporated at low temperature and pressure. Further drying i a vacuum desiccator left the sodium salt of ot-Z- thienyldiphenylmethylpenicillin as an almost White solid (6.94 g.).

The product inhibited Staph. aureus at a concentration of 0.5 mcg./ml., the benzylpenicillin-resistant Staph. 1 at 0.6 mcg./m1., and the benzylpenicillin-resistant Staph. 2 at 0.6 meg/ml.

a-2-thienyldip-henylacetyl chloride, M.P. 76-79 C., was prepared by heating the corresponding acid in benzene with thionyl-chloride and trace of pyridine.

(k) l-ARYLCYCLOALKYL TYPE Example 56.-1-phenylcyclopentylpenicillin A solution of 1-phenylcyclopentane-l-carbonyl chloride (6.0 g.) in dry acetone (30 ml.) Was'added gradually to a stirred solution of 6-aminopenicill'anic acid (5.63 g.) and sodium bicarbonate (6.55 g.) in Water ml.) and acetone (60 ml.). When addition was complete the mixture was stirred at room temperature for 2 /2 hours and then extracted with ether (2 x 90 ml.), only the aqueous phase being retained. This aqueous solution was covered with ether (50 ml.) and adjusted to pH 2 by the addition of N hydrochloric acid. After separating the layers, the aqueous phase was extracted with two 25 ml. portions of ether. The combined ether solutions (which at this stage contained the freepenicillin acid) were washed with water (50 ml.) and then shaken with enough 8% sodium bicarbonate solution to give a neutral aqueous phase (pH 7). The layers were separated and the ether phase was extracted with two 5 ml. portions of water to each of which was added sufiicient bicarbonate solution to produce an aqueous phase of pH 7. The combined aqueous solu- Example 57.1-phenylcyclolzexylpencilliit (l) 9-SUBSTITUTED-9-FLUORENYL TYPE Example 58.-9-p-meth0xyphenyl-9-fluorenylpenicillin A solution of 9-p-rnethoxyphenylfiuorene-9-carbonyl chloride (0.67 g.) in dry chloroform (8 ml.) was added dropwise to a stirred mixture of 6-aminopenicillanic acid (0.43 g.), chloroform (12 ml.), and triethylamine (0.84 ml.). Stirring was continued for 2 /2 hours and the mixture was then filtered. The filtrate was treated with successive small portions of 0.2 N hydrochloric acid, with vigorous shaking between additions, until the aqueous phase was at pH 3. The acid layer was separated and discarded, and the chloroform solution was washed with Water (20 ml.). Sufiicient 3% aqueous sodium bicarbonate solution was added to the chloroform solution to give, after vigorous shaking, a neutral emulsion (pH 7 This was evaporated at low temperature and pressure and the residual yellow solid dried in a Vacuum desiccator. There was obtained 0.83 g., of the sodium salt of 9-p-methoxyphenyl-9-fiuorenylpenicillin.

The product inhibited Staph. aureus at a concentration of 1.25 mcg./ml., and the benzylpenicillin-resistant Staph. 1 at 2.5 mcg./ml., and the benzylpenicillin-resistant Staph. 2 at 2.5 meg/ml. v

9-pmethoxyphenylfluorene-9-carbonyl chloride, M.P. 169171 C., was prepared by heating 9-p-methoxyphenylfluorene-9-carboxylic acid in benzene with thionyl chloride and a trace of pyridine.

' Example 59.9-methoxy-9-flu0renylpenicillin 9-methoxyfluorene-9-carboxylic acid (3.81 g.), thionyl chloride (1.1 ml), and pyridine (10 drops) in benzene ml.) were refluxedfor 75 minutes then evaporated in vacuo. The residual impure waxy acid chloride was dissolved in dry acetone ml.) and added dropwise to a stirred solution of 6-aminopenicilla11ic acid (3.42 g.) and sodium bicarbonate (4 g.) in water (48 ml.) and acetone (20 ml.). When addition was complete, the mixture was stirred at room temperature for 3 /2 hours, then diluted with water (50 ml.) and extracted with ether (2 x 50 ml.), only the aqueous phase being retained. This aqueous solution was covered with ether (50 ml.) and adjusted to pH 2.5-3 by the addition of N hydrochloric acid. After separating the layers, the aqueous phase was extracted with three further ml. portions of ether. The combined ether solutions (which at this stage contained the free penicillin acid) were washed with water and then shaken with sufficient 8% bicarbonate solution to give a neutral aqueous phase (pH 7). The layers were separated and the ether phase was extracted with two 5 ml. portions of water to each of which was added sufiicient bicarbonate solution to produce an aqueous phase of pH 7. Evaporation of the combined aqueous solutions at low temperature and pressure left the sodium salt of 9-methoxy-9-fiuorenylpenicillin, which was finally dried in a vacuum desiccator. Yield 3.2 g.

The product inhibited Staph. Oxford at a concentration of 0.25 meg/ml, Staph. 1 at'25 mcg./ml., and Staph. 2 at 12.5 rncg./ml.

Example 60.9-th0xy-9-flu0renylpenicillin 9-ethoxyfluorene-9-carboxylic acid (3.92 g.) was converted to the crude acid chloride, and the latter was used to acylate 6-aminopenicillanic acid (3.42 g.) as described in Example 59. The sodium salt of 9-ethoxy-9-fiuoreny1- penicillin was obtained as a white solid (5.63 g.).

It inhibited Staph. Oxford at a concentration of 1.25 mcg./m1.

Example 61 1- (4-nitrophenyl) -1-cyclohexanecarboxylic acid,

1- (4'-bromophenyl 1 -cyclopentanecarboxylic acid,

1- (4'-t-butylphenyl -9-fluorenecarboxylic acid,

1- (4-trifluoromethylphenyl -1-cyclohexanecarboxylic acid,

1- 3 '-aminopheny1)-l-cyclopentanecarboxylic acid,

1-(4'-sulfamylphenyl -1-(2'-methylcyclohexanc)- carboxylic acid,

1-(2'-benzylphenyl) -1-(2'-methylcyclohexane) carboxylic acid,

1- 3'-methoxyphenyl -9-fiuorenecarboxylic acid,

l-(2'-iodophenyl) -1-cyclohexanecarboxylic acid,

1- 3 '-diethylaminopheny1) -1-cyclopentanecarboxylic acid,

1- (4'-cyclohexylphenyl -9-fiuorenecarboxylic acid,

1- (4-methylaminophenyl) -1-cyclohexanecarboxylic acid,

1- (4'-chlorophenyl l-cyclopentanecarboxylic acid,

1- (2'-acetamidophenyl)-9-fluorenecarboxylic acid,

1- (3 ,4'-dimethylphenyl -1-cyclohcxanecarboxylic acid,

1-( 3',4'-dichloropheny1) -l-cyclopentanecarboxylic acid,

1-(3',4',5'-trirnethoxyphenyl) -9-fluorenecarboxylic acid,

1-(4'-hydroxyphenyl) -1-cyclohexanecarboxylic acid,

1- (3 ',4-dihydroxyphenyl -1-cyclopentanecarb oxylic acid,

1- 4'-methylthiophenyl -9-fl-uorenecarboxylic acid,

1- (4'methylsulfonylphenyl) -1-cyclohexanecarboxylic acid, and

1- 2',5 -dimethyl-4-hydroxyphenyl) -1-cyclopentacarb oxylic acid, respectively, to produce the acids 6-[1-(4'-nitrophenyl)-1-cyclohexanecarboxamido]- penicillanic acid,

6-[1-4'-bromophenyl)-1-cyclopentanecarboxamido] penicillanic acid,

6-[ 1-( 4'-t-butylphenyl) -9-fluorenecarboxamido] penicillanic acid,

6- 1-(4-trifluorornethylphenyl) -1-cyclohexanecarboxamido1penicillanic acid,

6- 1- 3 'arninop-henyl)-1-cyc10pentanecarb0Xa.mid0] penicillanic acid, I

6-[ l-(4'-sulfarnylphenyl) -1-(2-methylcyclohexane) carboxamido peenicillanic acid,

6-[1-(2-benzylphenyl)-1-(2'-methylcyclohexane)carbox amido] penicillanic acid,

6-[ 1- 3 -methoxyphenyl) -9-fiuorenecarboxamido] penicillanic acid,

6-[1-(2'-iodophenyl)-1-cyclohexanecarboxamido] penicillanic acid,

6- 1- 3 '-diethylarninopheny1)-1-cyc1opentanecarboxamido1penicillanic acid,

6-[1-(4'-cyclohexylphenyl)-9-fluorenecarboxamido] penicillanic acid,

6-[1-(4-.met hylaminophcnyl)-1-cyclohexanecarboxamido] penicillanic acid,

6-[ 1-(4-'-chloropheny1) -1-cyclopentanecarboxamido] penicillanic acid,

6-[ l- 2'-acetamidophenyl) -9-fluorenecarboxamido] penicillanic acid,

6- 1- (3 ',4'-dimethylphenyl -1-cyclohexanecarboX- a'rnido)penicil1anic acid,

(m) OTHER TYPES Example 62.Trichlrmethylpenicillin 6-aminopenicillanic acid (3.0 g.) was treated with trichloroaocetyl chloride (2.8 g.) as described in Example 1 to give the sodium salt of trichloromethylpenicillin as a pale yellow hygroscopic solid (1.7 g.). It inhibited Staph. aureus at a concentration of 125 mcg./ml.

Example 63.-a-Chl0r0-a-cyclohexylbenzylpenicillin A solution of a-chloro-a-cyclohexyl-u-phenylacetyl chloride (0.63 g.), in dry chloroform ml.) was added dropwise to a stirred mixture of 6-aminopenicillanic acid (0.43 g.), chloroform (12 ml.), and triethylamine (0.84 ml.). Stirring was continued for 2 /2 hrs, and the resulting solution was treated with successive small portions of 0.2 N hydrochloric acid, with vigorous shaking between additions, until the aqueous phase was at pH 2-3. The acid layer was separated and described,

and the chloroform solution was washed with water ml.). Sufiicient 3% aqueous sodium bicarbonate solution was added to the chloroform solution to give, after vigorous shaking, a neutral emulsion (pH 7). This was evaporated at low temperature and pressure and the residual yellow solid dried in a vacuum desiccator. There was obtained 0.84 g. of the sodium salt of a-chloro-a-cyclohexylbenzylpenicillin.

The product inhibited Staph. aareus at a concentration of 0.25 mcg./ml., Staph. Oxford at 0.025 mcg./ml., and the benzylpenicillin-resistant Staph. 1 at 12.5 mcg./ ml., and the benzylpenicillin-resistant Staph. 2. at meg/ml.

Example 64.a-Bromo-a-ethylpropylpenicillin A solution of a-bromo-u-ethylbutyryl bromide (7 g.) in dry acetone ml.) was added dropwise during 5 mins. to a stirred solution of 6-aminopenicillanic acid (5 .85 g.) and sodium bicarbonate (6.8 g.) in water (80 ml.) and acetone (40 ml.). When addition was complete the mixture was stirred at room temperature for 3 hrs., then diluted with water (40 ml.), and extracted with ether (2 x 80 ml.,), only the aqueous phase being retained. This aqueous solution was covered with ether ml.) and adjusted to pH 2 by the addition of N hydrochloric acid. After separating the layers, the aqueous phase was extracted with 25 ml. portions of ether. The combined ether solution (which at this stage contained the free penicillin acid) were washed with water (50 ml.) and then shaken with suflicient 8% sodium bicarbonate solution to give a neutral aqueous phase-(p-H 7). The layers were separated and the ether phase was extracted with two 5 ml. portions of water to each of which was added enough bicarbonate solution to produce an aqueous phase of pH 7. The combined aqueous solutions were washed with ether (15 ml.) and then evaporated at low temperature and pressure to leave the sodium salt of a-bromoa-ethylpropylpenicillin as a white solid, which was finally dried in a vacuum desiccator. Yield 9.26 g.

The product inhibited Staph. Oxford at a concentration of 0.12 mcg./ml., Staph. 1. at 25 mcg./ml., and Staph. 2. at 12.5 mc./ml.

The compounds of the present invention may be employed in admixture with suitable pharmaceutical carriers in various medicinal dosage forms. The present invention therefore also includes a composition comprising a pharmaceutical carrier and a compound of the general Formula II.

All temperatures herein are given in degrees Centigrade.

In summary, there are provided according to the present invention various 6-(a,a,a-tri-substituted-acetamido)penicillanic acids and their nontoxic salts which for ease of understanding may be divided into groups following the pattern used above, as follows:

(a) Triaryl type:

wherein R R and R are each members selected from the group consisting of hydrogen, nitro, amino, (lower)alkylamino, di-(lower)alkylamino, acylamino [where the acylating acid is an aliphatic carboxylic acid containing from one to ten carbon atoms inclusive and the substituent may thus also be named (lower)alkanoylamino], (lower) alkyl, fluoro, chloro, bromo, iodo, (lower)alkoxy, hydroxy, (lower)alkylthio, (lower)alkylsulfonyl, sulfamyl, benzyl, cyclohexyl, cyclopentyl and trifluoromethyl and R and R are each members selected from the group consisting of hydrogen, hydroxy, (lower) alkoxy and (lower)alkyl (including, both here and above, straight and branched chain saturated aliphatic groups having from one to ten carbon atoms inclusive).

(1)) Diarylalkyll type.

wherein R R and R are each members selected from the group consisting of hydrogen, nitro, amino, (lower)alkylamino, di-(lower)alkylamino, acylamino [where the acylating acid is an aliphatic carboxylic acid containing from one to ten carbon atoms inclusive and the substituent may thus also be named (lower) alkanoylamino], (lower) alkyl, fluoro, chloro, bromo, iodo, (lower)alkoxy, hydroxy, (lower)alkylthio, (lower)alkylsulfonyl, sulfamyl, benzyl, cyclohexyl, cyclopentyl and trifluoromethyl and R is a member selected from the group consisting of hydrogen, hydroxy, (lower) alkoxy and (lower)alkyl (including both here and above, straight and branched chain saturated aliphatic groups having from one to ten carbon atoms inclusive) and R represents a member selected from the group consisting of (lower)alkyl (that is, straight and branched chain saturated aliphatic groups having from one to ten carbon atoms inclusive) and cycloalkyl (e.g., cyclobutyl, cyclopentyl, cyclohexyl and cycloheptyl).

(c) Aryldialkyl type:

35 wherein R R and R are each members selected from the group consisting of hydrogen, nitro, amino, (lower)alkylamino, di-(lower)alkylamino, acylamino (where the acylating acid is an aliphatic carboxylic acid containing from one to ten carbon atoms inclusive and the substituent may thus also be named (lower) alkanoylamino), (lower)alkyl, fluoro, chloro, bromo, iodo, (lower)a1koxy, hydroxy, (lower) alkylthio, (lower)alkylsulfonyl, sulfamyl, benzyl, cyclohexyl, cyclopentyl and trifluoromethyl and R and R are each members selected from the group consisting of (lower)alkyl (including straight and branched chain saturated aliphatic groups having from one to ten carbon atoms inclusive) and cycloalkyl (e.g., cyclobutyl, cyclopentyl, cyclohexyl and wherein R R and R are each members selected from the group consisting of (lower)alkyl (including straight and branched chain saturated aliphatic groups having from one to ten carbon atoms inclusive) and cycloalkyl (e.g., cyclobutyl, cyclopentyl, cyclohexyl and cycloheptyl) (e) Diaryl(substituted oxy) type:

wherein R R and R are each members selected from the group consisting of hydrogen, nitro, amino, (lower)a1kylamino, di-(lower)alkylamino, acylamino [where the acylating acid is an aliphatic carboxylic acid containing from one to ten carbon atoms inclusive and the substituent may thus also be named (lower)a'lkanoylamino], (lower)alkyl, fiuoro, chloro, bromo, iodo, (lower)alkoxy, hydroxy, (lower)alkylthio, (lower) alkylsulfonyl, sulfamyl, benzyl, cyclohexyl, cyclopentyl and trifluoromethyl and R represents a member selected from the group consisting of -(lower)alkyl (that is, straight and branched chain saturated aliphatic groups having from one to ten carbon atoms inclusive) and cycloalkyl (e.g., cyclobutyl, cyclopentyl, cyclohexyl and cycloheptyl).

(f) DiaryKsubstituted thio) type! R1 l- R3 CH I II R45(|3CNH-CHCH C-CHa wherein R R and R are each members selected from the group consisting of hydrogen, nitro, amino, (lower)- alkylamino, di(lower)alkylamino, acylamin-o (where the acylating acid is an aliphatic carboxylic acid containing from one to ten carbon atoms inclusive .and the substituent may thus also be named (lower) al kanoylamino, (lower)- alkyl, fluoro, chloro, bromo, iodo, (lower)alkoxy, hydroxy, (lower)alkylthio, (lower)alkylsulfonyl, sulfamyl,

benzyl, cyclohexyl, cyclopentyl and trifluoromethyl and R represents a memiber selected from the group consisting of (lower)alkyl (that is, straight and branched chain saturated aliphatic groups having from one to ten carbon atoms inclusive) and cycloalkyl (e.g., cyclobutyl, cyclopentyl, cyc-lohexyl and cycloheptyl) and aralkyl (e.g., benzyl, B-phenethyl); and

wherein Ri, R and R are each members selected from the group consisting of hydrogen, nitro, amino, (lower)- alkylamino, di(lower)alkylamino, acylamino (where the acylating agent is an aliphatic carboxylic acid containing from one to ten carbon atoms inclusive and the substituent may thus also be named (lower) alkanoylamino) (lower)- alkyl, fluoro, chloro, bromo, iodo, (lower)alkoxy, hydroxy, (lower)alkylthio, (lower)alkylsulfonyl, sulfamyl, benzyl, cyclohexyl, cyclopentyl and trifluoromethyl and R is a member selected from the group consisting of hydrogen, hydroxy, (lower)alkoxy and (lower)alky1 (including, both here and above, straight and branched chain saturated aliphatic groups having from one to ten carbon atoms inclusive).

(g) Dialkyl (substituted oxy) type:

R2 in O=C--NCHCOOH 3 wherein R R and R are each members selected from the group consisting of hydrogen, nitro, amino, (lower)- alkylamino, di(lower) alkylamino, acylamino (Where the acylating acid is an aliphatic carboxylic acid containing from one to ten carbon atoms inclusive and the substituent may thus also be named (lower)alkanoylamino), (lower)alkyl, chloro, bromo, iodo, (lower)alkoxy, hydroxy, sulfamyl, benzyl, cyclohexyl, cyclopentyl, and trifluoromethyl, and R and R are each members selected from the group consisting of (lower)alkyl (including, both here and above, straight and branched chain saturated aliphatic groups having from one to ten carbon atoms inclusive), phenyl and phenyl(lower) alkyl (including benzyl, ozand p-phenethyl and 41-, pand 'y-phenylpropyl).

(h) Dialkyl(substituted thio) type:

wherein R R and R are each members selected from the group consisting of hydrogen, nitro, amino, (lower)- alkylamino, di(lower)all ylamino, a-cylamino (Where the acylating acid is an aliphatic carboxylic acid containing from one to ten carbon atoms inclusive and the substituent may thus also be named (lower)alkanoylamino), (lower)a-lkyl, fiuoro, chloro, bromo, iodo, (lower) alkoxy, hydroxy, (lower) alkylthio, (lower) alkylsulfonyl, sulfamyl, benzyl, cyclohexyl, cyclopentyl and trifluoromethyl and R and R are each members selected from the group consisting of (lower)alkyl (including straight and branched chain saturated aliphatic groups having from one to ten carbon atoms inclusive) and cycloalkyl (cg, cyclobutyl, cyclopentyl, cyclohexyl and cyclo'heptyl) wherein Ring A is .a member selected from the group consisting of cyclopentyl, (ilower) alkylcyclopentyl, cyclohexyl, (lower)alkylcyclohexyl and 9-fluorenyl and X represents a member selected from the group consisting of (lower) alkoxy and wherein R R and R are each members selected from the group consisting of hydrogen, nitro, amino, (lower)- alkylamino, di(lower) alkylamino, acylamino (where the acylating acid is an aliphatic carboxylic acid containing from one to ten carbon atoms inclusive and the substituent may thus also be named (lower) alkanoylamino), (lower) alkyl, fluoro, chloro, bromo, iodo, (lower) alkoxy, hydroxy, (lower)alkylthio, (lower) alkylsulfonyl, sulfamyl, benzyl, cyclohexyl, cyclopentyl and trifluoromet-hyl.

(k) Diaryl heterocyclic and miscellaneous types:

These types are exemplified by the following penicillins: a,a-diphenyl-2-thienylmethylpenicillin, 9-p-methoxypheny1-9-fiuorenylpenicillin, 9-methoxy-9-fluorenylpeniciliin, 9- ethoxy-9-fluorenylpenicillin, trichlormethylpenicillin, achloro-a-cyclohexylbenzylpencillin, a-bromo-a-ethylpropylpenicillin.

We claim:

1. A member selected from the group consisting of an acid having the formula wherein R R and R are each members selected from the group consisting of hydrogen, nitro, amino, (lower)- alkylamino, di(lower) alkylami-no, (lower) alkanoylamino, (lower)alkyl, chloro, bromo, iodo, (lower)alkoxy, hydroxy, sulfamyl, benzyl, cyclopentyl, cyclophexyl and trifluoromethyl, and R and R are each members selected from the group consisting of (=lower)alkyl, phenyl and phenyl(lower)alkyl; and their sodium, potassium, calcium, aluminum and ammonium salts and their nontoxic substituted ammonium salts with an amine selected from the group consisting of tri(lower)-alkylamines, procaine, dibenzylamine, N-benzyl-beta-phenethylamine, l-ephenamine, N,N'-dibenzylethylene diamine, dehydroabietyl amine and N,N bis-dehydroabietylethylenediamine.

2. A member selected from the group consisting of the acids having the formula R 0 CH3 O-CHs S I II --salts with an amine selected from the group consisting of tri(lower)alkylamines, procaine, dibenzylamine, N-

.benzyl-beta-phenethylamine, l-ephenamine, N,N-dibenzylethylenediamine, dehydroabietylamine and N,Nbisdehydroabietylethylenediamine.

38 3. A member selected from the group consisting of an acid having the formula wherein R R and R are each members selected from the group consisting of hydrogen, nitro, amino, (lower) alkylamino, di(lower) alkylamino, (lower) alkanoylamino, (lower)alkyl, fluoro, chloro, bromo, iodo, (lower)alkoxy, hydroxy, (lower)alkylthio, (lower)alkylsulfonyl, sulfamyl, benzyl, cyclohexyl, cyclopentyl and trifluoromethyl and R and R are each members selected from the group consisting of hydrogen, hydroxy, (lower)alkoxy and (lower)alkyl and its sodium, potassium, calcium and aluminum and ammonium salts and its nontoxic substituted ammonium salts with an amine selected from the group consisting of tri(lower)alkylamines, procaine, dibenzylamine, N-benZyl-beta-phenethylamine, 1- ephenamine, N,N'-dibenzylethylenediamine, dehydroabietylamine and N,Nbis-dehydroabietylethylenediamine.

4. A member selected from the group consisting of wherein R R and R are each members selected from the group consisting of hydrogen, nitro, amino, (lower) alkylamino, di-(lower)alkylamino, (lower) alkanoylamino, (lower)alkyl, fluoro, chloro, bromo, iodo, (lower) alkoxy, hydroxy, (lower)alkylthio, (lower)alkylsulfonyl, sul'famyl, benzyl, cyclohexyl, cyclopentyl and trifluoromethyl and R is a member selected from the group consisting of hydrogen, hydroxy, (lower)alkoxy and (lower) wherein R R and R are each members selected from the group consisting of hydrogen, nitro, amino, (lower) alkylamino, di-(lower) alkylamino, (lower) alkanoylami- 'no, (lower)alkyl, fluoro, chloro, bromo, iodo, (lower) alkoxy, hydroxy, (lower) alkylthio, (lower)alkylsulfonyl, sulfamyl, benzyl, cyclohexyl, cyclopentyl and trifluoromethyl and R and R are each members selected from the group consisting of (loWer)alkyl and cycloalkyl, and its sodium, potassium, calcium, aluminum and ammonium salts and its substituted ammonium salts with nontoxic amines selected from the group consisting of tri(lower) alkylamines, procaine, dibenzylamine, N-benzyl-beta- 39 phenethylamine, l-ephenamine, N,N'-dibenzylethylenediamine, dehydroabietylamine and N,N'-bis-dehydroabietylethylenediamine.

6. A member selected from the group consisting of an acid having the formula lh CH wherein R R and R are each members selected from the group consisting of (lower)alkyl and cycloalkyl, and its nontoxic sodium, potassium, calcium, aluminum and ammonium salts, and its nontoxic substituted ammonium salts with an amine selected from the group consisting of tri(lower) alkylamines, procaine, dibenzylamine, N- benzyl-beta-phenethylamine, l-ephenamine, N,N'-dibenzylethylenediamine, dehydroabietylamine and N,N-bisdehydroabietylethylenediamine.

7. A member selected from the group consisting of an acid having the formula 0 CH3 ll 8. A member selected from the group consisting of an R3 wherein R R and R are each members selected from the group consisting of hydrogen, nitro, amino, (lower)- alkylamino, di(lower) alkylamino, (lower)-alkanoylamino, (lower) alkyl, fluoro, chloro, bromo, iodo, (lower)-alkoxy, hydroxy, (lower) alkylthio, (lower)alkylsulfonyl, sulfamyl, benzyl, cyclohexyl, cyclopentyl .and trifluoromethyl and R represents a member selected from the group consisting of (loweryalkyl, cycloalkyl and aralkyl and its sodium, potassium, calcium, aluminum and ammonium salts and its nontoxic substituted ammonium salts, with amines selected from the group consisting of tri(lower) alkylamines, procaine, dibenzylamine, N-benzyl-betaphenethylamine, l-ephenamine, N,N-dibcnzylethylene- 40 diamine, dehydroabietylamine and- N,N'-bis-dehydroabietylethylenediamine.

9. A member selected acid having the formula from the group consisting of an wherein R R and R are each members selected from the group consisting of hydrogen, nitro, amino, (lower)- alkylarnino, di(lower) alkylamino, (lower) alkanoylamino, (lower):alkyl, fluoro, chloro, bromo, iodo, (lower) alkoxy, hydroxy, (lower)alkylthio, (lower)alkylsulfonyl, sulfamyl, benzyl, cyclohexyl, cyclopentyl and trifluoromethyl, and R is a member selected from the group consisting of hydrogen, hydroxy, (lower)a1koxy and (lower)alkyl, and'its sodium, potassium, calcium, aluminum and ammonium salts and its nontoxic substituted ammonium salts with amines selected from the group consisting of tri- (lower)alkylamines, procaine, dibenzylamine, N-benzylbetaphenethylamine, l-ephenamine, N,N-dibenzylethylenediamine, dehydroabietylamine and N,N-bis-dehydroabietylethylenediamine.

10. A member selected from acid having the formula the group consisting of an I l, s on, SC|l-C-NH--OH-Qfi o oHa Rt R4 0=(IJ-N--CHCOOH' wherein R R and R are each members selected from the group consisting of hydrogen, nitro, amino, (lower)- alkylamino, di(lower)-alkylamino, (lower)alkanoylamino, (lower) alkyl, fluoro, chloro, bromo, iodo, (loweflalkoxy, hydroxy, '(lower)alkylthio, '(lower)alkylsulfonyl, sulfamyl, benzyl, cyclohexyl, cyclopentyl and trifluoromethyl and R and R are each members selected from the group consisting of (lower) alkyl and cycloalkyl and its sodium, potassium, calcium, aluminum and ammonium salts and its substituted ammonium salts, with amines selected from the group consisting of tri(lower)alkylamines, procaine, dibenzyl-amine, N-benzyl-beta-phenethylamine, l-ephenamine, N,N'-dibenzylethylenediamine, dehydroabietyl amine and N,N-bis 1ehydroabietylethylenediamine.

11. A member selected from the group consisting of an acid having the formula wherein Ring A is a member selected from the group consisting of cyclopentyl, (lower) alkylcyclopentyl, cyclohexyl, (lower)alkylcyclohexyl and 9-fluorenyl and X represents a member selected from the group consisting of (lower)alkoxy and 3 wherein R R and R are each members selected from the group consisting of hydrogen, nitro, amino, (lower)- alkylamino, di(lower) alkylamino, (lower) alkanoylamino, (lower) alkyl, fluoro, chloro, bromo, iodo, (lower)a1koxy, hydroxy, (lower)alkylthio, (lower):alkylsulfonyl, sulfamyl, benzyl, cyclohexyl, cyclopentyl, and trifluorometh- 

1. A MEMBER SELECTED FROM THE GROUP CONSISTING OF AN ACID HAVING THE FORMULA
 3. A MEMBER SELECTED FROM THE GROUP CONSISTING OF AN ACID HAVING THE FORMULA
 8. A MEMBER-SELECTED FROM THE GROUP CONSISTING OF AN ACID HAVING THE FORMULA
 11. A MEMBER SELECTED FROM THE GROUP CONSISTING OF AN ACID HAVING THE FORMULA 